Neurological Disorders

Coding Dimension ID: 
303
Coding Dimension path name: 
Neurological Disorders

Antagonizing aging and senescence of myocardial stem cells with Pim-1 kinase

Funding Type: 
Basic Biology II
Grant Number: 
RB2-01602
ICOC Funds Committed: 
$0
Disease Focus: 
Epilepsy
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Public Abstract: 
In the last several decades, average lifespan has increased significantly. More individuals reach 85-90 years of age but often suffer from health related problems with severe limitations in functional activity, underscoring the need to improve the health and quality of life of this patient population. An additional critical question is whether the current limit in life expectancy actually reflects the ineluctable genetic clock or the increased incidence of congestive heart failure (CHF) with aging has interfered with the programmed death of the organ and organism, negatively influencing quality of life and maximum lifespan in humans. Morbidity and mortality for CHF continue to increase and parallel the extension in median lifespan of the population, pointing to aging as the major risk factor of the human disease. Recent data published from the American Heart Association document that there are 5.7 million patients affected by this disease in the United States alone with an incidence of 670,000 new cases per year. Importantly, the overwhelming majority of individuals with CHF are 65 years of age or older. The aging heart typically shows a decreased functional reserve and limited capacity to adapt to abrupt changes in hemodynamic load. But the critical question is whether the senescent heart is the product of aging-associated events or the result of a primary aging effect on the number and function of cardiac progenitor cells (CPC). A recent study strengthened the notion that myocyte replacement occurs in the human heart at a rate higher in young adults than in individuals at 75 years of age. CPCs and myocytes undergo replicative senescence with severe telomere shortening and expression of proteins typically found in old cells. These molecular modifications dictate irreversible growth arrest and cell death activation. Senescent CPCs and poorly contracting markedly hypertrophied myocytes accumulate, the pool of functionally competent CPCs is reduced, and cardiac decompensation supervenes defining the senescent heart phenotype. Although myocardial aging and heart failure may not necessarily deplete myocardial CPCs, strategies capable of interfering with telomere attrition, cellular senescence and death have three important objectives: a) Preventing and/or delaying the exhaustion of the CPC compartment; b) Enhancing the viability, engraftment and growth of the delivered cells within the tissue; and c) Increasing the efficiency of CPCs which, in turn, reduces dramatically the number of cells to be administered and, therefore, the time between cell isolation/expansion and treatment. Thus, the long-term objective is to identify whether genetic engineering of CPC with survival and proliferative signals promotes a youthful cardiac phenotype characterized by increased myocyte regeneration and preservation / expansion of the CPC pool.
Statement of Benefit to California: 
In 2004, the total healthcare cost in the US was1.9 trillion dollars and cardiovascular disease ranked as the most costly disease category, accounting for 8.3% of these overall costs. Cardiovascular disease and, in particular, myocardial infarction resulting in congestive heart failure, continues to be the number one cause of death in the US, killing some 300,000 Americans each year. Congestive heart failure is also the leading cause of hospital admissions. There are nearly 5 million Americans who are suffering from this illness, with 550,000 new cases reported annually. Despite recent advances in drug therapies for acute myocardial infarction, the average five-year survival for patients suffering this condition remains only at roughly 50% level. While cardiac transplantation is a well-established treatment for end-stage congestive heart failure, this treatment is limited to only 2,000 patients per year due to a severe and chronic shortage of acceptable donor hearts. As the most populous state, California bears the greatest public health impact of myocardial infarction and congestive heart failure as well, a burden that is only getting heavier as the population ages. There is a clear need for novel therapies for these disease conditions, the development of which will benefit California tremendously, from its population to the state’s public health system and its foundation in biomedical research. One such cutting edge treatment uses the genetic engineering technology which has already been shown to much promise in relevant animal models and is on track for translational implementation in the clinical setting. In addition to the cost saving benefits that will be realized by the California healthcare system as a result of the development of this technology, there will also be significant economic development benefits realized by the State. All the funds will be reinvested in California, fueling economic growth and recovery in addition to furthering a likely revolutionary treatment of heart failure. This funding will also serve to create new jobs associated with the development of genetic modification of progenitor cells to enhance their regenerative capacity. Synergy can result as other cell therapies using related technologies and targeting different disease areas are likely to develop as a direct consequence of this funding, assuming a positive outcome for the current project. These funds will also directly support education and jobs at {REDACTED}, which are undergoing significant and painful budget reductions and losing critical talent.
Progress Report: 
  • We have been developing new methods to identify the products of stems cells that are differentiated in tissue culture dished. We are focusing on generating a specific type of neuron - cortical interneuron. To this end, we have identified specific sequences in the human genome that drive gene expression in the immature cortical interneurons. Results from the first year of our work provide evidence that our method to use these gene expression elements is working to help us identify cortical interneurons.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders, such as epilepsy.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders.

Pioneer Factor Interactions at Tissue-Specific Enhancers in Pluripotent Cells

Funding Type: 
Basic Biology II
Grant Number: 
RB2-01602
ICOC Funds Committed: 
$0
Disease Focus: 
Epilepsy
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Public Abstract: 
Embryonic stem cells (ESC) hold great promise for the treatment of debilitating diseases and for use as research tools to elucidate the molecular mechanisms underlying many diseases. Current knowledge is already quite advanced and has allowed researchers to begin making use of stem cells for clinical applications. However, the full clinical potential of ESC cannot be realized without a better understanding of their fundamental properties. Increased fundamental knowledge is necessary to help overcome technical limitations in our ability to differentiate ESC into clinically useful cell types and to enhance the efficiency and consistency with which differentiated cells obtained from patients in need of stem cell-based therapies can be reprogrammed to an ESC-like state. ESC lines are of great value because of their unique ability to continually divide on laboratory tissue-culture dishes while maintaining pluripotency, which is defined as a capacity to differentiate into almost any cell type. Pluripotency is known to be dependent on the expression of key genes whose protein products function by turning on many other genes that are needed to achieve the pluripotent state. Considerable attention has also been focused on genes encoding proteins that regulate the differentiation of ESC into specific cell lineages. These genes possess intriguing properties that keep them silent in ESC, but poised for activation when the ESC receive appropriate differentiation signals. The research proposed in this application focuses on a third class of genes that have received relatively little attention from researchers studying pluripotency: genes expressed only in differentiated cell types, which generally remain silent until long after the ESC have differentiated into a specific cell lineage. Although prior models suggested that the cascade of events leading to the activation of these typical tissue-specific genes does not begin until the ESC have differentiated, recent evidence from our laboratory and others supports a hypothesis in which the competence of these genes for expression in differentiated cells is dependent on specific marks established at their DNA regulatory regions in the pluripotent state. The proposed examination of this hypothesis will reveal whether the proper marking of tissue-specific genes in ESC is essential for their differentiation into clinical useful cell types and tissues and for the proper functioning of mature ESC-derived tissues.
Statement of Benefit to California: 
The research proposed in this application will increase our knowledge of the fundamental properties of embryonic stem cells (ESC) that are important for their pluripotency, defined as their capacity to differentiate into almost all human cell types and tissues. ESC hold great promise for treating or leading to a better understanding of human diseases, which would greatly benefit the State of California and its citizens. However, despite this potential and the rapid progress that has been made toward its fulfillment, our incomplete knowledge of the properties of ESC critical for their pluripotency has limited their utility. Previous studies of the key determinants of pluripotency have focused on genes that are actively expressed in ESC or that play a role in the initial decision to differentiate into specific cell lineages. The research proposed in this application focuses on an emerging hypothesis that pluripotency also requires the active marking of typical tissue-specific genes, which generally remain silent until long after the ESC have begun to differentiate. The proper marking of these genes in ESC may be essential for their competence for activation in differentiated tissues. A rigorous evaluation of this hypothesis will contribute to efforts to identify the sources of variability in the differentiation potential of ESC lines and to develop improved methods for the reprogramming of differentiated cells to a pluripotent state.
Progress Report: 
  • We have been developing new methods to identify the products of stems cells that are differentiated in tissue culture dished. We are focusing on generating a specific type of neuron - cortical interneuron. To this end, we have identified specific sequences in the human genome that drive gene expression in the immature cortical interneurons. Results from the first year of our work provide evidence that our method to use these gene expression elements is working to help us identify cortical interneurons.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders, such as epilepsy.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders.

Basic Mechanisms Underlying Human Cardiac Cell-Cell Junction Differentiation in Human iPSC

Funding Type: 
Basic Biology II
Grant Number: 
RB2-01602
ICOC Funds Committed: 
$0
Disease Focus: 
Epilepsy
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Public Abstract: 
Heart disease is the number one cause of death and disability in California and in the United States. Especially devastating is Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC), an inherited form of heart disease associated with a high frequency of arrhythmias and sudden cardiac death in young people. It is a common heart disease in young athletes, who despite their appearance of health are struck down by this type of heart disease. Understanding this disease is key. Even though it is an inherited disease early detection is hindered because parents carrying the genetic code have highly variable clinical symptoms, making ARVC and catastrophic cardiac events very hard to predict and avoid. There is some evidence that this kind of heart disease is caused by mistakes in the genetic code essential for holding the mechanical integrity of heart muscle cells together or cell junctions. What is missing is an understanding of the basic biology of these heart muscle cell junctions in humans and appropriate human model systems to study their biology and dynamics in the absence and presence of disease. The goal of the studies proposed here is to understand the basic biology of how the human heart muscle cell junctions differentiate and mature and what happens when normal development goes wrong. We will do this by deriving and studying heart muscle cells from normal populations derived from human induced pluripotent stem cells (iPS) and heart muscle cells derived from human iPS from people we know are carrying mistakes in the genetic code for cell junction components. Human iPS are a unique population of stem cells from our own tissues, such as skin, that have the same genetic information as the rest of our bodies. Therefore, human iPS from people who carry the ARVC heart disease mistakes can be used in our laboratory to provide a true human model of that disease. Recent advances in stem cell biology have highlighted the unique potential of human iPS that could be used in the future as a source of cells for cell-based therapies for heart disease. However, before there can be any clinical application of these approaches, we need a detailed fundamental understanding of the basic biology and differentiation/maturation of these human iPS into the heart muscle cells. In this proposal we also propose to determine whether the microenvironment can influence human heart muscle cell junction differentiation/maturation in the absence and presence of disease in the human iPS model system, so that we might be able to obtain high yielding cultures of fully mature functional cells, a future goal for realizing stem cell medicine for heart disease. Knowledge gained from these studies will advance the field by providing us with model systems to study the biology of human cardiac muscle cell junction differentiation/maturation, which are crucially important in understanding human cell-cell junction diseases such as ARVC.
Statement of Benefit to California: 
Heart disease is the number one cause of death and disability within the United States and the rates are calculated to be even higher for citizens of the State of California when compared to the rest of the nation. These diseases place tremendous financial burdens on the people and communities of California, which highlights the need to find therapies to alleviate these growing burdens. Especially devastating is Arrhythmogenic Right Ventricular Cardiomyopathy (ARVC), an inherited form of heart disease associated with a high frequency of arrhythmias and sudden cardiac death in young people. Even though it is an inherited disease early detection is hindered because parents carrying the genetic code have highly variable clinical symptoms, making ARVC and catastrophic cardiac events very hard to predict and avoid. There is some evidence that this kind of heart disease is caused by mistakes in the genetic code essential for holding the mechanical integrity of heart muscle cells together or cell junctions. What is missing is an understanding of the basic biology of these heart muscle cell junctions in humans and appropriate human model systems to study their biology and dynamics in the absence and presence of disease. The goal of the studies proposed here is to understand the basic biology of how the human heart muscle cell junctions differentiate and mature and what happens when normal development goes wrong using human induced pluripotent stem cells (iPS) as a model system. Recent important advances in stem cell biology have highlighted the unique potential of human iPS, which is an endogenous population of stem cells, which can be derived from our own tissues, such as skin, that could be used in the future as a source of cells for patient-specific cell-based therapies for heart disease. However, before there can be any clinical application of these approaches, a fundamental understanding of the basic biology and maturation/differentiation of these unique stem cells into heart muscle cells and muscle cell-cell junctions will be required. Our proposal meets this goal and tries to identify avenues to advance stem cell medicine for heart disease by developing model systems to systematically characterize and manipulate the maturation and differentiation of human iPS into fully mature heart muscle cells by studying the basic biology behind how heart muscle cell junctions differentiate and mature. Since various forms of human heart diseases, including ARVC, have underlying defects in muscle cell junctions, this knowledge will be crucially important in unraveling the biological defects burdening Californians with these forms of heart disease alongside advancing the application of stem cell based approaches as therapies for heart disease, such as ARVC, for citizens of the State of California.
Progress Report: 
  • We have been developing new methods to identify the products of stems cells that are differentiated in tissue culture dished. We are focusing on generating a specific type of neuron - cortical interneuron. To this end, we have identified specific sequences in the human genome that drive gene expression in the immature cortical interneurons. Results from the first year of our work provide evidence that our method to use these gene expression elements is working to help us identify cortical interneurons.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders, such as epilepsy.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders.

Molecular regulation of cell survival and cellular interactions of pluripotent stem cells

Funding Type: 
Basic Biology II
Grant Number: 
RB2-01602
ICOC Funds Committed: 
$0
Disease Focus: 
Epilepsy
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Public Abstract: 
The recent technological breakthrough has paved the way to convert adult differentiated cells such as skin cells into undifferentiated cells called human induced pluripotent stem (hiPS) cells that have an ability (pluripotency) to become all types of adult cells in human body. Because of this enormous ability, hiPS cells are thought to be the potential source of the cell-transplantation therapy for the treatment of diseases such as Parkinson's disease and diabetes mellitus. Despite this promising function of hiPS cells, due to its short history, there are many biological questions and practical hurdles to overcome. One questions is how we can distinguish fully converted genuine iPS cells from incompletely converted cells. Other important issues include development of technologies to derive and grow hiPS cells at high efficiency under completely animal-free conditions for future medical purposes. We have recently identified a signaling pathway (like a hormone that mediates biological information) that controls cell-cell attachment of pluripotent stem cells. Moreover, by controlling this pathway, we have established a method to grow another type of pluripotent cells, human embryonic stem (hES) cells, in a fully animal-free condition. In this proposal, we will investigate whether measuring the activity of this pathway could be helpful to select genuine iPS cells. Furthermore, by using this technology, we will develop a new method to efficiently generate and expand a new hiPS cell lines that are completely free from animal-derived materials.
Statement of Benefit to California: 
Our research will focus on developing novel technologies by which hES and iPS cells can be propagated under completely defined conditions. The establishment of such a new method would impact virtually all hES and iPS cell-based application programs as it involves a common basic process to expand undifferentiated pluripotent stem cells before turning into any type of adult cells for the therapeutic purposes. It is therefore predictable that the new methodology will be promptly translated as an intellectual property to be commercialized, and would substantially activate the biotechnology field in the State of California. More importantly, the new methodology will be provided to the Institutes in California at the highest priority where the method will accelerate the process to apply the pluripotent cell-based transplantation approach for the clinical settings that would further substantiate the enhancement of the medical environment for California citizens.
Progress Report: 
  • We have been developing new methods to identify the products of stems cells that are differentiated in tissue culture dished. We are focusing on generating a specific type of neuron - cortical interneuron. To this end, we have identified specific sequences in the human genome that drive gene expression in the immature cortical interneurons. Results from the first year of our work provide evidence that our method to use these gene expression elements is working to help us identify cortical interneurons.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders, such as epilepsy.
  • We have identified 5 gene regulatory elements (enhancers) that can promote gene expression in a specific type of neuronal precursor and neuron. We found that these enhancers can be used to aid in the identification and isolation of these types of cells from embryonic stem cells. In other studies, our group is testing the feasibility of using these types of cells to ameliorate neurological disorders.

Autologous Delivery of Pim-1 Enhanced Cardiac Stem Cells: A Novel Clinical Therapy for Cardiac Muscle Regeneration Post MI

Funding Type: 
Disease Team Research I
Grant Number: 
DR1-01480
ICOC Funds Committed: 
$0
Disease Focus: 
Stroke
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
Public Abstract: 
There have been consistent advancements in the prevention and treatment of cardiovascular disease. Despite these advancements and partly due to the increasing age of the human population, heart attacks continue to be a human plague currently affecting 5 million people. Heart attack is the most common cause of hospital admission leading to 300,000 deaths each year in the US. The average 5-year survival for these patients is only 50%. The most aggressive treatment for these patients is heart transplantation but this is limited to only 2,000 patients each year because of a serious lack of suitable donors. Therefore, there is an urgent need for new ways to treat patients suffering from heart failure. In the last 10 years, much work has been done on the use of stem cell therapy for organ failure. Recent reports indicate some successes using stem cell therapies to treat heart attack but only limited, short-term benefit has been seen. {REDACTED} has discovered a new type of stem cell therapy for heart attack. This technique uses stem cells taken from the patient’s own heart. The cells, called adult human cardiac progenitor cells (hCPCs), are modified to produce a protein (Pim-1) known to protect the heart and also found to protect hCPCs after they are returned to the patient’s heart. Animal data shows that Pim-1 modified hCPCs are able to generate new heart tissue that persists for at least 6 months and causes major improvements in the heart’s ability to work. In this proposal, the {REDACTED} will partner with {REDACTED} and his colleagues at {REDACTED}, who are recognized experts in tracking fate of stem cells. They will validate survival of Pim-1 modified hCPCS in small and large animal models using state-of-the-art molecular imaging technologies. The third member of the disease team is {REDACTED}. {REDACTED} exists specifically to advance the Pim-1 technology to the point of evaluation by the Food and Drug Administration (FDA) for potential use in human patients. We anticipate that the work conducted in this proposal will result in an application to FDA for the first human clinical trial using the patient’s own hCPCs, modified to produce Pim-1, as a therapy for heart attack and prevention of subsequent heart failure through the generation of new heart muscle. Funding of this proposal will: 1) Develop procedures for Pim-1 modification in hCPCs, 2) Develop quality-assured procedures and perform clinical quality manufacturing of Pim-1 modified hCPCs, 3) Demonstrate safety and efficacy of these modified hCPCs under quality-assured pre-clinical conditions in large animal models, 4) Design the human clinical trial protocol and related documentation that will be proposed to FDA, 5) Compile all documents required for the submission of the Investigational New Drug Application (IND) to FDA, and 6) Submit the IND to FDA by the end of year 4 of this proposal.
Statement of Benefit to California: 
In 2004, the total healthcare cost in the US was 1.9 trillion dollars and cardiovascular disease ranked as the most costly disease category, accounting for 8.3% of these overall costs. Cardiovascular disease and, in particular, myocardial infarction resulting in congestive heart failure, continues to be the number one cause of death in the US, killing some 300,000 Americans each year. Congestive heart failure is also the leading cause of hospital admissions. There are nearly 5 million Americans who are suffering from this illness, with 550,000 new cases reported annually. Despite recent advances in drug therapies for acute myocardial infarction, the average five-year survival for patients suffering this condition remains only at roughly 50% level. While cardiac transplantation is a well-established treatment for end-stage congestive heart failure, this treatment is limited to only 2,000 patients per year due to a severe and chronic shortage of acceptable donor hearts. As the most populous state, California bears the greatest public health impact of myocardial infarction and congestive heart failure as well, a burden that is only getting heavier as the population ages. Thus, there is a clear need for development of novel therapies for these disease conditions, the development of which will benefit California tremendously, from its population to the state’s public health system and its foundation in biomedical research. One such cutting edge treatment uses the Pim-1 modified cardiac progenitor cell (CPC) technology developed by investigators at {REDACTED}, which has already shown significant promise in relevant animal models of ischemic heart disease. In addition to the cost saving benefits that will be realized by the California healthcare system as a result of the development of this technology, there will also be significant economic development benefits realized by the State. Although biotechnology is a robust business sector in California, this sector is currently suffering from the economic downturn as venture funding dries up and jobs are being lost. Funding of this proposal will directly support existing jobs within California’s biotechnology industry at the firms designated for the development work. All the funds specifically targeting scientific procedures will be reinvested in California, fueling economic growth and recovery in addition to furthering a likely revolutionary treatment of heart failure. This funding will also serve to create new jobs at {REDACTED} and other companies associated with the Pim-1 modified hCPC technology. Synergy can result as other cell therapies using related technologies and targeting other disease areas are likely to develop as a consequence of this funding. These funds will also directly support education and jobs at {REDACTED} coping with painful budget reductions resulting in loss of critical talent and research momentum.
Progress Report: 
  • A stroke kills brain cells by interrupting blood flow. The most common “ischemic stroke” is due to blockage in blood flow from a clot or narrowing in an artery. Brain cells deprived of oxygen can die within minutes. The loss of physical and mental functions after stroke is often permanent and includes loss of movement, or motor, control. Stroke is the number one cause of disability, the second leading cause of dementia, and the third leading cause of death in adults. Lack of movement or motor control leads to job loss and withdrawal from pre-stroke community interactions in most patients and institutionalization in up to one-third of stroke victims. The most effective treatment for stroke, thrombolytics or “clot-busters”, can be administered only within 4.5 hours of the onset of stroke. This narrow time window severely limits the number of stroke victims that may benefit from this treatment. This proposal develops a new therapy for stroke based on embryonic stem cells. Because our (and others’) laboratory research has shown that stem cells can augment the brain’s natural repair processes after stroke, these cells widen the stroke treatment opportunity by targeting the restorative or recovery phase (weeks or months after stroke instead of several hours).
  • Embryonic stem cells can grow in a culture dish, but have the ability to produce any tissue in the body. We have developed a technique that allows us to restrict the potential of embryonic stem cells to producing cell types that are found in the brain, making them “neural stem cells”. These are more appropriate for treating stroke and may have lower potential for forming tumors. When these neural stem cells are transplanted into the brains of mice or rats one week after a stroke, the animals are able to regain strength in their limbs. Based on these findings, we propose in this grant to further develop these neural stem cells into a clinical development program for stroke in humans at the end of this grant period.
  • A multidisciplinary team is working rigorously to test the effectiveness of stem cell delivery in several models of stroke, while simultaneously developing processes for the precise manufacture, testing and regulatory approval of a stem cell therapy intended for human use. Each step in this process consists of definite milestones that are being achieved, providing measurable assessment of progress toward therapy development. To accomplish this task, the team consists of stroke physician/scientists, pharmacologists, toxicologists, experts in FDA regulatory approval and key collaborations with a biotechnology manufacturer active in this area. This California-based team has a track record of close interactions and brings prior stroke clinical trial and basic science experience to the proposed translation of a stem cell therapy for stroke.
  • In the first year of this program, the cells have been translated from an encouraging research level to a product which can be manufactured under conditions suitable for human administration. This has included optimization of the production process, development of reliable tests to confirm cell identity and function, and characterization of the cells utilizing these tests. In animal models in two additional laboratories , improvement in motor function following stroke has been confirmed. The method of administration has also been carefully studied. It has been determined that the cells will be administered around the area of stroke injury rather than directly into the middle of the stroke area. These results encourage the translation of this product from research into clinical trials for the treatment of motor deficit following stroke.
  • A stroke kills brain cells by interrupting blood flow. The most common “ischemic stroke” is due to blockage in blood flow from a clot or narrowing in an artery. Brain cells deprived of oxygen can die within minutes. The loss of physical and mental functions after stroke is often permanent and includes loss of movement, or motor, control. Stroke is the number one cause of disability, the second leading cause of dementia, and the third leading cause of death in adults. Lack of movement or motor control leads to job loss and withdrawal from pre-stroke community interactions in most patients and institutionalization in up to one-third of stroke victims. The most effective treatment for stroke, thrombolytics or “clot-busters”, can be administered only within 4.5 hours of the onset of stroke. This narrow time window severely limits the number of stroke victims that may benefit from this treatment. This proposal develops a new therapy for stroke based on embryonic stem cells. Because our (and others’) laboratory research has shown that stem cells can augment the brain’s natural repair processes after stroke, these cells widen the stroke treatment opportunity by targeting the restorative or recovery phase (weeks or months after stroke instead of several hours).
  • Embryonic stem cells can grow in a culture dish, but have the ability to produce any tissue in the body. We have developed a technique that allows us to restrict the potential of embryonic stem cells to producing cell types that are found in the brain, making them “neural stem cells”. These are more appropriate for treating stroke and may have lower potential for forming tumors. When these neural stem cells are transplanted into the brains of mice or rats one week after a stroke, the animals are able to regain strength in their limbs. Based on these findings, we propose in this grant to further develop these neural stem cells into a clinical development program for stroke in humans at the end
  • of this grant period.
  • A multidisciplinary team is working rigorously to test the effectiveness of stem cell delivery in several models of stroke, while simultaneously developing processes for the precise manufacture, testing and regulatory approval of a stem cell therapy intended for human use. Each step in this process consists
  • of definite milestones that are being achieved, providing measurable assessment of progress toward therapy development. To accomplish this task, the team consists of stroke physician/scientists, pharmacologists, toxicologists, experts in FDA regulatory approval and key collaborations with a biotechnology manufacturer active in this area. This California-based team has a track record of close interactions and brings prior stroke clinical trial and basic science experience to the proposed translation of a stem cell therapy for stroke.
  • A stroke kills brain cells by interrupting blood flow. The most common “ischemic stroke” is due to blockage in blood flow from a clot or narrowing in an artery. Brain cells deprived of oxygen can die within minutes. The loss of physical and mental functions after stroke is often permanent and includes loss of movement, or motor control. Stroke is the number one cause of disability, the second leading cause of dementia, and the third leading cause of death in adults. Lack of movement or motor control leads to job loss and withdrawal from pre-stroke community interactions in most patients and institutionalization in up to one-third of stroke victims. The most effective treatment for stroke, thrombolytics or “clot-busters”, can be administered only within 4.5 hours of the onset of stroke. This narrow time window severely limits the number of stroke victims that may benefit from this treatment. This proposal develops a new therapy for stroke based on embryonic stem cells. Because our (and others’) laboratory research has shown that stem cells can augment the brain’s natural repair processes after stroke, these cells widen the stroke treatment opportunity by targeting the restorative or recovery phase (weeks or months after stroke instead of several hours).
  • Embryonic stem cells can grow in a culture dish, but have the ability to produce any tissue in the body. We have developed a technique that allows us to restrict the potential of embryonic stem cells to producing cell types that are found in the brain, making them “neural stem cells”. These are more appropriate for treating stroke and may have lower potential for forming tumors. When these neural stem cells are transplanted into the brains of mice or rats one week after a stroke, the animals are able to regain strength in their limbs. Based on these findings this grant is supporting conduct of IND-enabling work to initiate a clinical development program for stroke in humans by the end of this grant period.
  • A multidisciplinary team is working rigorously to test the effectiveness of stem cell delivery in several models of stroke, while enabling precise manufacture, testing and regulatory clearance of a first in human clinical trial. Defined milestones are being achieved, providing measurable assessment of progress toward therapy development. Definitive manufacturing and pharmacology studies are underway and regulatory filings are in progress. The team consists of stroke physician/scientists, pharmacologists, toxicologists, experts in FDA regulatory and key collaborations with a biotechnology manufacturer active in this area. This California-based team has a track record of close interactions and brings prior stroke clinical trial and basic science experience to the proposed translation of a stem cell therapy for stroke.
  • A stroke kills brain cells by interrupting blood flow. The most common 'ischemic stroke' is due to blockage in blood flow from a clot or narrowing in an artery. Brain cells deprived of oxygen can die within minutes. The loss of physical and mental functions after stroke is often permanent and includes loss of movement, or motor, control. Stroke is the number one cause of disability, the second leading cause of dementia, and the third leading cause of death in adults. Lack of movement or motor control leads to job loss and withdrawal from pre-stroke community interactions in most patients and institutionalization in up to one-third of stroke victims. The most effective treatment for stroke, thrombolytics or 'clot-busters', can be administered only within 4.5 hours of the onset of stroke. This narrow time window severely limits the number of stroke victims that may benefit from this treatment. This proposal develops a new therapy for stroke based on embryonic stem cells. Because our (and others') laboratory research has shown that stem cells can augment the brain's natural repair processes after stroke, these cells widen the stroke treatment opportunity by targeting the restorative or recovery phase (weeks or months after stroke instead of several hours).
  • Embryonic stem cells can grow in a culture dish, but have the ability to produce any tissue in the body. We have developed a technique that allows us to restrict the potential of embryonic stem cells to producing cell types that are found in the brain, making them 'neural stem cells'. These are more appropriate for treating stroke and may have lower potential for forming tumors. When these neural stem cells are transplanted into the brains of mice or rats one week after a stroke, the animals are able to regain strength in their limbs. Based on these findings this grant is supporting conduct of IND-enabling work to initiate a clinical development program for stroke in humans by the end of this grant period.
  • A multidisciplinary team is working to test the effectiveness of stem cell delivery in several models of stroke, while enabling precise manufacture, testing and regulatory clearance of a first in human clinical trial. Defined milestones are being achieved, providing measurable assessment of progress toward therapy development. Definitive manufacturing and pharmacology studies are underway and regulatory filings are in progress. The team consists of stroke physicians/scientists, pharmacologists, toxicologists, experts in FDA regulatory and key collaborations with a biotechnology manufacturer active in this area. This California-based team has a track record of close interactions and brings prior stroke clinical trial and basic science experience to the proposed translation of a stem cell therapy for stroke.

Hematopoietic Stem Cell Transplants for Severe Combined Immune Deficiency and Systemic Sclerosis

Funding Type: 
Disease Team Research I
Grant Number: 
DR1-01471
ICOC Funds Committed: 
$0
Disease Focus: 
Amyotrophic Lateral Sclerosis
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
Public Abstract: 
Blood stem cells, which reside in the bone marrow (BM) can generate every type of blood and immune cell. They are the only cells necessary to re-establish blood formation if the BM is wiped out by disease or by treatments such as radiation or chemotherapy, as is the case for people who undergo a BM transplant. BM transplants have been performed for >50 years as life-saving procedures for many illnesses. However, patients do not receive pure blood stem cells, and the procedure is considered high risk mainly because BM cells received from a donor contain a combination of blood stem cells plus other mature immune cells. These mature cells pose a conundrum to physicians because on the one hand, the donor’s mature cells can be beneficial to the patient by assisting blood stem cells to take root and grow in the recipient as well as potentially helping battle tumors in cancer patients. However on the other hand, these mature donor cells can attack the recipient’s tissues, perceiving them as foreign and causing a syndrome called graft-versus-host disease (GVHD). Unfortunately, 10-20% of patients that undergo a transplant die from the consequences of GVHD. In the last decade technologies were developed to purify blood stem cells eliminating mature immune cells, thereby eliminating the danger of GVHD. However, transplant physicians remained hesitant to use such grafts because of concerns that purified stem cells without the accompanying immune cells would not take and grow in the recipient. Members of this team have therefore worked out new ways in mice that may be used on patients so that they will accept purified blood stem cell grafts without significant side effects. The reagents we will develop belong to a class of proteins called antibodies. The specialized antibodies we will use are biologic tools that allow us to both purify human stem cells, and eliminate blood stem cells in the recipient thereby clearing the BM for donor cells. We plan to adapt the technologies that have worked successfully in mice to treat two different disorders for whom BM transplant can be curative, but if performed by conventional methods is very high risk and can be fatal for the patient. The disorders we aim to cure by this approach are the childhood disease called severe combined immune deficiency (SCID), and the other is an autoimmune disease called systemic sclerosis (SSc). Children born with SCID lack immune cells to fight infections and without treatment die within the first year of life. Patients with severe forms of SSc experience thickening and tightening of the skin, lung and gastrointestinal problems which ultimately results in death after several years of suffering. We intend for these studies to result in superior treatments for these diseases. Since blood stem cell transplants have the capability of curing many other childhood and autoimmune disease, the ultimate impact of our studies will potentially be on a much broader spectrum of diseases.
Statement of Benefit to California: 
In 2004 California citizens passed a historic proposition supporting research that could result in the use of stem cells to cure many diseases. As a result, public and private institutions in California have emerged as leaders in this field, and scientists are now well on the path to producing tissues from primitive embryonic stem cells (ESCs). As scientists learn to direct these cells to become the tissues needed to replace damaged or failing ones, the obstacle of a patient rejecting these new tissues is a problem that must be overcome. The studies proposed by this Team address this issue. Tissues or organs are rejected because they come from donors who are genetically different. Similarly, tissues derived from ESCs will be genetically different from patients who need these tissues and therefore at risk for rejection. In order to prevent tissue rejection, patients that undergo transplants of organs (i.e, heart, kidney, lung) must remain life-long on medications to suppress their white blood cells from rejecting the grafts. There is one group of transplant patients that are routinely taken off their immune suppressive drugs -- bone marrow transplant (BMT) patients. These patients undergo BMT to cure them of severe cancers or inherited blood diseases. However, they can be liberated from their immune suppressive drugs because donor blood forming stem cells that take root in their bodies make the white blood cells that decide which tissues are identified as “foreign” or “self”. New white blood cells re-educate the recipient’s immune system to accept donor tissues as self. Thus, a state of harmony called immune tolerance is achieved so that donor blood is made without difficulty, and, in theory, the recipient can accept transplanted organs from the marrow donor without need for immune suppression. A similar strategy can be adapted to induce immune tolerance to tissues derived from ESCs. Remarkably, BMT also has the capability to cure autoimmune diseases such as multiple sclerosis, juvenile diabetes and many others. The major obstacle to use BMT beyond the treatment cancers has been the dangers associated with the procedure. This Team will take a crucial step to make BMT safer by transplanting only purified blood stem cells. The benefits of these potential advancements to our state are many. First and foremost is the health and well-being of all Californians who face the many diseases treatable by BMT. In addition, it is a simple fact that with every major scientific advancement come immediate economic benefits to the region that generated those advancements. These benefits can manifest in the form of academic donations from sources around the world, service industries that support the medical establishments that practice the procedures, and hi tech companies who receive their funding globally. This activity can all result in greater investment in California and continued job creation that has made California such a desirable place to live.
Progress Report: 
  • Considerable progress was made on transitioning cells and cell production methods from research-scale to translational/clinical scale. Specifically, Year 1 activities were focused on transitioning from research to pilot-scale cell production methods, and characterization of the animal amyotrophic lateral sclerosis (ALS) disease model. These activities were essential because cellular therapy development is a multi-stage process with increasing stringency over time in terms of the increased focus on the details of the methods, stringent requirements for reagents/materials, greater scale, and more thorough product characterization during the transition from early research to an approved cellular therapy.
  • During Year 1, small-scale embryonic stem cell (ESC) growth and differentiation methods previously developed for research at Life Technologies were further developed at a larger pilot-scale, which provided enough cells to perform early animal pre-clinical studies and cell characterization. In addition to the increased scale of cell production, where possible, research grade reagents and materials were substituted with reagents and materials that would be required or preferred for producing a cell therapy for use in humans [produced under Good Manufacturing Practices (GMP), non-animal origin, well characterized]. These conditions are not ideal for many ESC lines, and only 1 of the 4 starting ESC lines was able to adapt successfully to these culture conditions. To increase the number of potential clinical ESC candidate cell lines, we acquired 2 additional ESC lines, UCFB6 and UCSFB7 from the University of California, San Francisco. Development is ongoing to ensure the cell processing methods are robust and scalable for the increased cell numbers required for the large-scale animal studies in Year 2. Cells from the pilot-scale production are being subjected to deep sequencing as part of the development of molecular characterization methods that may provide future quality control assays.
  • During Year 1, further studies of a rat ALS disease model were performed to: 1) optimize cell injection methods; 2) improve characterization of disease onset and progression in the rat model; 3) evaluate the utility of behavioral and electrophysiology tests for following the disease; and 4) evaluate histology methods for measuring neuron damage and detection of implanted cells, which will be used to optimize the large-scale efficacy studies planned for Year 2. We discovered that several time-consuming analysis approaches for efficacy evaluation could be replaced by simpler, more cost effective approaches. Additionally, the Year 1 studies tested and ensured that the team could handle an aggressive cell implant schedule, twice daily immunosuppression, demanding behavioral and electrophysiology assessments, and extensive histology evaluations.
  • Considerable progress was made on transitioning cells and cell production methods from research-scale to translational/clinical scale, including initial cell production in a GMP facility with GMP compatible production methods. Additionally, extensive characterization of the amyotrophic lateral sclerosis (ALS) disease animal model was completed and cells were evaluated for potential efficacy in this ALS disease animal model. These activities are key for continued progress in cellular therapy development, which is a multi-stage process that requires increasing focus on the details of the methods, stringent requirements for reagents/materials, greater scale, and more thorough product characterization during the transition to an approved cellular therapy.
  • Specifically, we made significant progress in three major areas:
  • First, we found evidence for efficacy using neural stem cells made at Life Technologies. In brief, during Year 1, the rat ALS disease model was shown to be a more aggressive disease model with an earlier disease onset and more rapid progression to end-stage and death than the model that had been used in previous studies. During Year 2, this more aggressive ALS disease model was further characterized with the identification of a reliable marker of disease onset, and demonstration that alpha motor neuron sparing by implanted cells could be detected and measured even, despite the aggressive nature of disease progression in this rat model.
  • We found that H9 NSCs produced by Life Technologies, when implanted into the rat ALS disease model, survived, migrated extensively into the area where alpha motor neurons are located, differentiated into cells that appear to be astrocytes, and provided a protective effect for the alpha motor neurons. This protective effect was determined by comparing the survival of alpha motor neurons on the side of the rat spinal cord where NSCs were implanted with the side of the spinal cord that did not have cells implanted. The side of the spinal cord where the NSCs were implanted showed approximately 10% more surviving alpha motor neurons than the matching side of the spinal cord that did not have cells implanted.
  • Second, cells from the various production methods were subjected to gene sequencing as part of the development of molecular characterization methods. This sequencing information was critical to identify whether cells produced by various methods were typical for the cell type, or exhibited qualities that indicated they were not optimal cell populations. These methods will be used to identify optimal markers for characterizing cell populations as part of current cell production development and for future quality control assays.
  • Third, during Year 2, Life Technologies further developed their pilot-scale embryonic stem cell (ESC) growth and differentiation methods to be more easily adaptable to cell production under Good Manufacturing Practices (GMP). This involved increasing the scale of cell production, and where possible, substituting reagent grade reagents and materials with reagents and materials that would be required or preferred for producing a cell therapy for use in humans (produced GMP, non-animal origin, well characterized). These conditions are not ideal for many ESC lines, and in Year 1, only one (H9) of the 4 starting ESC lines was successfully adapted to these culture conditions, however, 3 additional ESC lines were acquired to increase the number of potential clinical ESC candidate cell lines. One of these ESC lines (UCSFB7 from the University of California, San Francisco) was successfully adapted to the pilot ESC culture conditions, and resulted in the production of NSCs, and with AP production in progress. Because the research version of ESC line H9 has been used to successfully produce NSCs at Life Technologies, agreements are in progress for City of Hope for NSC cell production using the H9 ESCs, that have been banked under GMP conditions at City of Hope. In addition, pilot-scale cell production was initiated earlier than originally planned at the University of California, Davis GMP facility. The plan is to produce NSCs and APs under conditions that UC Davis has found to be successful in the past, and transition these methods to GMP compliance. To date, UC Davis has produced ESCs from 3 ESC lines [UCSF4, UCSF4.2 (a.k.a. UCSFB6) and UCSF4.3 (a.k.a. UCSFB7] and has produced NSCs from ESC line UCSF4. The UCSF4 NSCs are scheduled to be shipped to UCSD for testing in the ALS disease animal model in early June, 2012, and NSC production from ESC lines UCSF4.2 and UCSF4.3 is expected to begin in late June 2012.

Mechanism of BCL6-dependent stem cell maintenance in B cell lineage leukemia

Funding Type: 
Basic Biology I
Grant Number: 
RB1-01358
ICOC Funds Committed: 
$0
Disease Focus: 
Parkinson's Disease
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Public Abstract: 
Despite significant advances in the treatment of leukemia over the past four decades, the rate of long-term survival has reached a plateau and still large numbers of leukemia patients die, mostly because of relapse and drug-resistance, which was recently attributed to the persistence of leukemia stem cells. If a therapy succeeds in eradicating leukemia stem cells, de novo initiation of the disease (relapse) is no longer possible. Therapeutic progress in recent clinical trials has likely been stalled, partly because current cytotoxic therapy approaches target proliferating bulk leukemia cells rather than quiescent leukemia stem cells. We now discovered that BCL6, a factor known to play a central role in B cell lymphomas, also plays a key role in the maintenance of leukemia stem cells. Since leukemia stem cells represent the origin of relapse and drug-resistance in leukemia in many cases, the identification of BCL6 as a target for leukemia stem cell eradication holds great promise. BCL6 is a master regulatory factor that controls the production of many different important genes. BCL6 was not previously known to be involved in leukemias. In preliminary studies for this proposal, we have discovered aberrant expression of BCL6 as a central component of a fundamentally novel pathway of leukemia stem cell self-renewal and drug-resistance in a wide array of human leukemias, some of which are still difficult to treat. In these leukemias, drug-treatment results in aberrant production of BCL6 by the leukemia cells, which appears to allow leukemia stem cell to self-renew and become resistance against drugtreatment. Recently a drug has been developed that can attach to BCL6 and block its cancer-causing activities. We found that this BCL6 inhibitor, which is called RI-BPI, has strong synergistic activity when combined with conventional drug-treatment, which opens up a powerful new therapeutic strategy for leukemia stem cell eradication through targeted inhibition of BCL6. Based on the discovery of BCL6 as a key component of a novel pathway of drug-resistance and stem cell self-renewal in a wide array of leukemias, we propose three Aims to develop these findings towards application in patient care: (1) To test the hypothesis that aberrant expression of BCL6 in human leukemia cells promotes leukemia stem cell survival, (2) To determine the frequency and phenotype of BCL6-dependent leukemia stem cells in human B cell ALL and (3) To validate a the role of the BCL6 inhibitor RI-BPI as a therapy for targeted eradication of leukemia stem cells. Since RI-BPI is currently going through the process of approval for use in clinical trials, we expect to be able to test the power of this approach in clinical trials by the end of the funding period.
Statement of Benefit to California: 
B cell lineage acute lymphoblastic leukemia (ALL) represents the most frequent malignancy in childhood and is frequent in adults as well. Thousands of children and adults in California are afflicted with B cell leukemia and a significant portion of these patients will ultimately die despite the tremendous progress that has been made in leukemia treatment. Compelling evidence indicates that many leukemias are not curable because currently available chemotherapy target the bulk of the rapidly dividing leukemia cells but not the rare drug resistant leukemia stem cells that are quiescent and do not divide. For this reason, current research efforts both by laboratory investigators and clinicians focus on the leukemia stem cells because they are widely considered as the origin of drug-resistance and recurrence of the disease. Ground-breaking research in other subtypes of leukemia that are more frequent in adults has recently identified the leukemia stem cell. Additional research even found the "Achilles heel" of these resilient leukemia cells and potential targets for future drug-therapy. Unfortunately, leukemia stem cells have not been identified in B cell lineage ALL, nor do we know the mechanism that enables drug-resistance in these leukemia stem cells. The absence of this information represents a major unsolved problem, because knowledge about the biology of the leukemia stem cells is required for the development of future drug-therapies that will help to eradicate leukemia stem cells in this frequent leukemia subset. This seems particularly important, since B cell lineage ALL accounts for about 30% of all childhood cancers and is by far the most frequent malignancy in children and teenagers. In summary, the benefits to the citizens of California from the CIRM disease specific grant in leukemia are: (1) direct benefit to the thousands of patients with B cell lineage leukemia (2) higher quality of life due to definitive and targeted treatments that avoid severe side-effects and long-term disabilities (3) new partnership between the laboratory and clinical investigators as well as the pharmaceutical sector in California that leads to new synergies (4) the deployment of a multidisciplinary approach for pathwayspecific drug-therapy that will be applicable to other types of cancer (5) Realization of the CIRM mandate to develop highly effective novel therapies within a short time to benefit the health of Californians suffering from B cell lineage ALL.
Progress Report: 
  • Parkinson's disease results primarily from the loss of neurons deep in the middle part of the brain (the midbrain), in particular neurons that produce dopamine (referred to as “dopaminergic”). In this region of the midbrain there are actually two different groups of dopaminergic (DA) neurons, and only one of them, the neurons of the substantia nigra (SN) are highly susceptible to degeneration in patients with PD. There is a relative sparing of the second group of midbrain dopaminergic neurons, called the ventral tegmental area (VTA) dopaminergic neurons. These two groups of neurons reside close to each other in the brain and both make dopamine. They are virtually indistinguishable except for one major functional difference—they release dopamine, the transmitter that is lost in Parkinson’s patients, to their downstream neuronal targets in different ways. SN neurons deliver dopamine in small rapid squirts, like a sprinkler, whereas VTA neurons have a tap that provides a continuous stream of dopamine.
  • A major therapeutic strategy for patients with PD is to make new DA neurons from human embryonic stem cells (hES). As stem cells have the potential to develop into any type of cell in the body, these considerations suggest that we should devise a way to produce SN neurons in the absence of VTA neurons from stem cells for use in transplantation. At present although we can produce dopaminergic neurons from hES cells, the scientific community cannot distinguish SN from VTA neurons in vitro due to lack of molecular markers or a bioassay, and we are therefore unable to identify culture conditions that favor the production of one over the other,
  • In addition to releasing dopamine differently, SN and VTA neurons have axons that project to different regions of the striatum. It has been shown over the last decade that specific classes of guidance cues guide axons to their particular targets. One approach we have taken has been to investigate whether differences in axon guidance receptor expression and or responses to guidance cues in vitro might provide both markers and a bioassay that will distinguish SN from VTA neurons. Over the last year we have shown that VTA and SN neurons respond differentially to Netrin-1 and express different markers associated with the guidance cue family. We now have a bioassay and markers that distinguish these two populations of neurons in vitro and in the coming year we plan to utilize this information to identify cultures conditions that favor the production of SN over VTA neurons, from hES cells.
  • Parkinson’s disease results primarily from the loss of neurons deep in the middle part of the brain (the midbrain), in particular neurons that produce dopamine (referred to as “dopaminergic”). In this region of the midbrain there are actually two different groups of dopaminergic (DA) neurons, and only one of them, the neurons of the substantia nigra (SN) are highly susceptible to degeneration in patients with PD. There is a relative sparing of the second group of midbrain dopaminergic neurons, called the ventral tegmental area (VTA) dopaminergic neurons. These two groups of neurons reside close to each other in the brain and both make dopamine. They are virtually indistinguishable except for one major functional difference—they release dopamine, the transmitter that is lost in Parkinson’s patients, to their downstream neuronal targets in different ways. SN neurons deliver dopamine in small rapid squirts, like a sprinkler, whereas VTA neurons have a tap that provides a continuous stream of dopamine. 
A major therapeutic strategy for patients with PD is to make new DA neurons from human embryonic stem cells (hES). As stem cells have the potential to develop into any type of cell in the body, these considerations suggest that we should devise a way to produce SN neurons in the absence of VTA neurons from stem cells for use in transplantation. At present although we can produce dopaminergic neurons from hES cells, the scientific community cannot distinguish SN from VTA neurons in vitro due to lack of molecular markers or a bioassay, and we are therefore unable to identify culture conditions that favor the production of one over the other, 
In addition to releasing dopamine differently, SN and VTA neurons have axons that project to different regions of the striatum. It has been shown over the last decade that specific classes of guidance cues guide axons to their particular targets. One approach we have taken has been to investigate whether differences in axon guidance receptor expression and or responses to guidance cues in vitro might provide both markers and a bioassay that will distinguish SN from VTA neurons. We showed previously that VTA and SN neurons respond differentially to Netrin-1 and express different markers associated with the guidance cue family. Also, in this year using backlabeling, laser capture and microarray analysis of SN vs VTA neurons, we have identified a number of genes expressed in on or the other population. We now have a bioassay and markers that distinguish these two populations of neurons in vitro and in the coming year we plan to utilize this information to identify cultures conditions that favor the production of SN over VTA neurons, from hES cells.
  • Parkinson's disease (PD) is a neurodegenerative movement disorder that affects more than six million people worldwide. The main symptoms of the disease result from the loss of neurons from the midbrain that produce dopamine (referred to as "dopaminergic" or DA neurons).Human embryonic stem cells (hESC) offer an exciting opportunity to treat Parkinson’s disease by transplanting hESC-derived DA neurons to replace those that have died. There are actually two groups of midbrain DA neurons in the human brain. Those from the substantia nigra (SN) are highly susceptible to degeneration in Parkinson's patients while those from the ventral tegmental area (VTA) are not. These two types of neurons have similar features but have different functions and it is important to ensure that DA neurons from hESC are the correct SN type before they are used in therapy. The primary goal of this research was to study these two neuronal types in animals and determine if the distinguishing features discovered in mice or rats can be used to more easily recognize and purify SN-type DA neurons made from hESC.
  • One of the discoveries made in this research is that SN and VTA neurons show differences in how they make connections within the brain. We have been able to identify some of the molecules that guide each neuron to connect to it appropriate target and have found that SN and VTA neurons placed in the petri dish can be distinguished from each other by their response to guidance molecules. Work in the final period of this grant has focused on testing guidance response in hESC-derived DA neurons and we have found that many of the neurons produced from hESC do show SN-like responses to guidance molecules. This discovery is being further developed as a screening tool to help guide our ongoing efforts to make increasingly pure populations of DA neurons from hESC.
  • Future human trials will likely utilize such DA neurons but since embryonic stem cells have the potential to develop into any type of cell in the body, it is important to ensure that the production methods used to make a therapeutic product for Parkinson’s disease do indeed specifically produce SN neurons. Prior to the research supported under this CIRM grant, the scientific community was not able to distinguish SN from VTA neurons outside of their normal brain environment and therefore had no ability to confirm whether a method produced one type selectively and not the other. Further refinements of the assay tools developed in our research may provide a practical means of quantifying the purity of a DA neuron preparation. This would have a significant impact transplantation therapy as well as provide useful insights into the molecular mechanisms that underlie proper connectivity and function of SN and VTA DA neurons in humans.

In vitro reprogramming of mouse and human somatic cells to an embryonic state

Funding Type: 
New Faculty I
Grant Number: 
RN1-00564
ICOC Funds Committed: 
$2 229 427
Disease Focus: 
Rett's Syndrome
Neurological Disorders
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 
Embryonic stem (ES) cells are remarkable cells in that they can replicate themselves indefinitely and have the potential to turn into all possible cell type of the body under appropriate environmental conditions. These characteristics make ES cells a unique tool to study development in the culture dish and put them at center stage for regenerative medicine. Two techniques, one called somatic cell nuclear transfer (SCNT) and the other in vitro reprogramming, have shown that adult cells from the mouse can be reverted to an ES like state. In SCNT, adult cell nuclei are transferred into oocytes and allowed to develop as early embryos from which ES cells can be derived, while in the in vitro method four genes are ectopically activated in the adult cell nucleus to induce an embryonic state in the culture dish. Key requirement for both processes is to erase the memory of the adult cell that specifies it as an adult cell and set up the ES cell program. How this happens remains unclear, and if it can be reproduced with human adult cells is an open question. Therefore, we will attempt to use the in vitro reprogramming method to generate human ES cells from adult cells and begin to understand the mechanism of the reprogramming process in both human and mouse cells. In addition to being integral to improving our understanding of how ES cells develop, if successful, this work will provide an important milestone for regenerative medicine. Many debilitating diseases and conditions are caused by damage to cells and tissue. In vitro reprogramming could provide a way to generate patient-specific stem cells that, in culture, could be turned into the type of cell or tissue needed to cure the patient’s disease or injury and transplanted back into the patient’s body. For example, Parkinson’s disease is caused by the loss or destruction of nerve cells. If reprogramming becomes possible, we could take a skin biopsy from a patient with Parkinson’s disease, induce the embryonic state in those skin cells to then be able to turn them into nerve cells and transplant them back into the same donor patient. Reprogramming could also be used to repair spinal cord injuries, allowing people who are paralyzed by accidents to walk again, or be helpful for patients with juvenile diabetes. One important advantage of patient-specific self-transplants is that they obviate the need for immunosuppression, which is often problematic for the patient. In addition, human cell reprogramming could be a new way to study how diseases progress at the cellular level as reprogramming could generate ES cells from patients with complex diseases that can be studied in detail for what makes them go awry during development. This knowledge could speed the search for new treatments and possibly cures for some of the most complex diseases that affect societies. We hope that the knowledge gained from our studies on reprogramming can, someday, support research that will help to put these idea to clinical use.
Statement of Benefit to California: 
Donated organs and tissues are often used to replace those that are diseased or destroyed, but unfortunately, the number of people needing a transplant exceeds the number of organs available for transplantation. Embryonic stem (ES) cells can be propagated in the laboratory for an unlimited period of time and can turn into all the specialized cell types that make us a human being. Therefore, ES cells offer the possibility of a renewable source of replacement cells and tissues to treat diseases, conditions, and disabilities such as Parkinson’s and Alzheimer’s, spinal cord injury, stroke, burns, heart disease, diabetes, osteoarthritis and rheumatoid arthritis. Our research is aimed to generate ES cells from adult cells through a method called in vitro reprogramming and to understand the mechanism by which the ES cell program can be reinstated in the adult cells. This work will not only provide the foundation for a better understanding of how human ES cells develop, but, if successful, be an important milestone for regenerative medicine. The advantage of using ES cells derived from adult cells by in vitro reprogramming would be that the patient’s own cells could be reprogrammed to an ES cell state and therefore, when transplanted back into the patient, not be attacked and destroyed by the body’s immune system. This would be beneficial to the people of California as tens of millions of Americans suffer from diseases and injuries that could benefit from research of in vitro reprogramming. Such advances would benefit the health as well as the economy of the state of California.
Progress Report: 
  • The discovery of induced pluripotent stem (iPS) cells by Shinya Yamanaka in 2006 marks a major landmark in the fields of stem cell biology and regenerative medicine. iPS cells can be obtained by co‐expression of four transcription factors in differentiated cells. The reprogramming process takes 2‐3 weeks and is very inefficient with about 1 in a 1000 somatic cells giving rise to an iPS cell. In previous work, we and others had demonstrated that mouse iPS cells are highly similar to ES cells in their molecular and functional characteristics as they for example can support adult chimerism with germline
  • contribution. The goal of the New Faculty Award proposal is to understand the molecular mechanisms underlying transcription factor‐ induced reprogramming of differentiated cells and to define the iPS cell state.
  • During this funding period, our efforts have focused on all three Aims. Within Aim 1, we have addressed a range of technical strategies to improve the reprogramming process. In Aim 2, we have analyzed human and mouse iPS cells in comparison to ES cells and attempted a better definition of the iPS cell state. In Aims 3, we are currently attempting to define barriers of the reprogramming process and begin to understand the transcriptional network that leads to reprogrammed cells.
  • The discovery of induced pluripotent stem (iPS) cells, which are derived from differentiated cells by simply overexpression a few transcription factors, by Shinya Yamanaka in 2006 marks a major landmark in the fields of stem cell biology and regenerative medicine. To unfold the full potential of reprogramming for disease studies and regenerative medicine, we believe that it is important to understand the molecular mechanisms underlying transcription factor‐ induced reprogramming and to carefully characterize the iPS cell state. To this end, during the third year of funding, we have devised a novel screen to identify factors important for the reprogramming process and allow replacement of the original reprogramming factors. We also studied the role of candidate transcriptional and chromatin regulators in the reprogramming process, which led us to identify novel barriers of the reprogramming process and to a better understanding of how chromatin interferes with the reprogramming process. We have also made progress in understanding the function of the reprogramming factors. Regarding human iPS cell lines, we have derived iPS cells from patients carrying X-linked diseases, and are beginning to characterize them molecularly. Together, we hope that our work will contribute to a better understanding of the reprogramming process.
  • Cellular reprogramming and the generation of induced pluripotent stem cells (iPSCs) from differentiated cells has enabled the creation of patient-specific stem cells for use in disease modeling. Reprogramming to the induced pluripotent state can be achieved through the ectopic expression of Oct4, Sox2, Klf4 and cMyc. Insight into the role that the reprogramming factors, various signaling pathways and epigenetic mechanisms play during the different stages of reprogramming remains limited, partly due to the low efficiency with which somatic cells convert to pluripotency. During the past year we have made great progress in (i) defining the molecular requirement for the reprogramming factors; (ii) gaining a better understanding of how repressive chromatin states control the reprogramming process; (iii) determining the differential regulation of chromatin states during reprogramming; (iv) identifying novel reprogramming stages; (v) assessing the three-dimensional organization of the genome during reprogramming; and (vi) determining the influence of a specific signaling pathway and its downstream effectors on different stages of the reprogramming process. Together, our findings provide novel mechanistic insights into the reprogramming process, which will form the basis of approaches to approve the efficiency of the process.
  • When this grant was awarded in 2008, reprogramming to the induced pluripotent state was just achieved by Shinya Yamanaka through the ectopic expression of Oct4, Sox2, Klf4 and cMyc in mouse fibroblasts. The overall goal of this proposal was to understand the molecular mechanisms underlying in vitro reprogramming of somatic cells of the mouse to iPSCs and to apply this knowledge to the reprogramming of human somatic cells. During the last funding period, our work particularly aimed at mechanistic questions: (i) determining the molecular origin of the spatio-temporal demarcation of the DNA binding sites of the reprogramming factors, and how the reprogramming factors induce chromatin changes, employing systematic and comprehensive mapping approaches; (ii) defining how the reprogramming factors induce a specific transcriptional output on target genes; (iii) identifying the steps of the reprogramming process to mouse iPSCs, which revealed an unprecedented detail of the reprogramming process and established that transition through a multitude of hierarchical stages is a fundamental feature of the reprogramming process; (iv) determining the dynamics of DNA methylation in reprogramming; (v) gaining a better understanding of how repressive Polycomb proteins control the reprogramming process; (vi) assessing the three-dimensional organization of the genome during reprogramming; and (vii) using the human iPSC approach for disease studies. Together, our findings provide novel mechanistic insights into the reprogramming process.

Banking transplant ready dopaminergic neurons using a scalable process

Funding Type: 
Early Translational II
Grant Number: 
TR2-01856
ICOC Funds Committed: 
$6 016 624
Disease Focus: 
Parkinson's Disease
Neurological Disorders
Collaborative Funder: 
Maryland
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 
Parkinson's disease (PD) is a devastating movement disorder caused by the death of dopaminergic neurons (a type of nerve cells in the central nervous system) present in the midbrain. These neurons secrete dopamine (a signaling molecule) and are a critical component of the motor circuit that ensures movements are smooth and coordinated. All current treatments attempt to overcome the loss of these neurons by either replacing the lost dopamine, or modulating other parts of the circuit to balance this loss or attempting to halt or delay the loss of dopaminergic neurons. Cell replacement therapy (that is, transplantation of dopaminergic neurons into the brain to replace lost cells and restore function) as proposed in this application attempts to use cells as small pumps of dopamine that will be secreted locally and in a regulated way, and will therefore avoid the complications of other modes of treatment. Indeed, cell therapy using fetal tissue-derived cells have been shown to be successful in multiple transplant studies. Work in the field has been limited however, partially due to the limited availability of cells for transplantation (e.g., 6-10 fetuses of 6-10 weeks post-conception are required for a single patient). We believe that human embryonic stem cells (hESCs) may offer a potentially unlimited source of the right kind of cell required for cell replacement therapy. Work in our laboratories and in others has allowed us to develop a process of directing hESC differentiation into dopaminergic neurons. To move forward stem cell-based therapy development it is important to develop scale-up GMP-compatible process of generating therapeutically relevant cells (dopaminergic neurons in this case). The overall goal of this proposal is to develop a hESC-based therapeutic candidate (dopaminergic neurons) by developing enabling reagents/tools/processes that will allow us to translate our efforts into clinical use. We have used PD as a model but throughout the application have focused on generalized enabling tools. The tools, reagents and processes we will develop in this project will allow us to move towards translational therapy and establish processes that could be applied to future IND-enabling projects. In addition, the processes we will develop would be of benefit to the CIRM community.
Statement of Benefit to California: 
Parkinson’s disease affects more than a million patients United States with a large fraction being present in California. California, which is the home of the Parkinson’s Institute and several Parkinson’s related foundations and patient advocacy groups, has been at the forefront of this research and a large number of California based scientists supported by these foundations and CIRM have contributed to significant breakthroughs in this field. In this application we and our collaborators in California aim propose to develop a hESC-based therapeutic candidate (dopaminergic neurons) that will allow us to move towards translational therapy and establish processes that could be applied to future IND-enabling projects for this currently non-curable disorder. We believe that this proposal includes the basic elements that are required for the translation of basic research to clinical research. We believe these experiments not only provide a blueprint for moving Parkinson’s disease towards the clinic for people suffering with the disorder but also a generalized blueprint for the development of stem cell therapy for multiple neurological disorders including motor neuron diseases and spinal cord injury. The tools and reagents that we develop will be made widely available to Californian researchers. We expect that the money expended on this research will benefit the Californian research community and the tools and reagents we develop will help accelerate the research of our colleagues in both California and worldwide.
Progress Report: 
  • Parkinson's disease (PD) is a devastating movement disorder caused by the death of dopaminergic neurons (a type of nerve cells in the central nervous system) present in the midbrain. These neurons secrete dopamine (a signaling molecule) and are a critical component of the motor circuit that ensures movements are smooth and coordinated.
  • All current treatments attempt to overcome the loss of these neurons by either replacing the lost dopamine, or modulating other parts of the circuit to balance this loss or attempting to halt or delay the loss of dopaminergic neurons. Cell replacement therapy (that is, transplantation of dopaminergic neurons into the brain to replace lost cells and restore function) as proposed in this application attempts to use cells as small pumps of dopamine that will be secreted locally and in a regulated way, and will therefore avoid the complications of other modes of treatment. Indeed, cell therapy using fetal tissue-derived cells have been shown to be successful in multiple transplant studies. Work in the field has been limited however, partially due to the limited availability of cells for transplantation (e.g., 6-10 fetuses of 6-10 weeks post-conception are required for a single patient).
  • We believe that human embryonic stem cells (hESCs) may offer a potentially unlimited source of the right kind of cell required for cell replacement therapy. Work in our laboratories and in others has allowed us to develop a process of directing hESC differentiation into dopaminergic neurons. To move forward stem cell-based therapy development it is important to develop scale-up GMP-compatible process of generating therapeutically relevant cells (dopaminergic neurons in this case).
  • The overall goal of this proposal is to develop a hESC-based therapeutic candidate (dopaminergic neurons) by developing enabling reagents/tools/processes that will allow us to translate our efforts into clinical use. We have used PD as a model but throughout the application have focused on generalized enabling tools. The tools, reagents and processes we will develop in this project will allow us to move towards translational therapy and establish processes that could be applied to future IND-enabling projects. In addition, the processes we will develop would be of benefit to the CIRM community.
  • Parkinson's disease (PD) is a devastating movement disorder caused by the death of dopaminergic neurons (a type of nerve cells in the central nervous system) present in the midbrain. These neurons secrete dopamine (a signaling molecule) and are a critical component of the motor circuit that ensures movements are smooth and coordinated.
  • All current treatments attempt to overcome the loss of these neurons by either replacing the lost dopamine, or modulating other parts of the circuit to balance this loss or attempting to halt or delay the loss of dopaminergic neurons. Cell replacement therapy (that is, transplantation of dopaminergic neurons into the brain to replace lost cells and restore function) as proposed in this application attempts to use cells as small pumps of dopamine that will be secreted locally and in a regulated way, and will therefore avoid the complications of other modes of treatment. Indeed, cell therapy using fetal tissue-derived cells have been shown to be successful in multiple transplant studies. Work in the field has been limited however, partially due to the limited availability of cells for transplantation (e.g., 6-10 fetuses of 6-10 weeks post-conception are required for a single patient).
  • We believe that human pluripotent stem cells (PSC) may offer a potentially unlimited source of the right kind of cell required for cell replacement therapy. Work in our laboratories and in others has allowed us to develop a process of directing PSC differentiation into dopaminergic neurons. To move forward stem cell-based therapy development it is important to develop scale-up GMP-compatible process of generating therapeutically relevant cells (dopaminergic neurons in this case).
  • During this grant, we have optimized a step-wise scalable process for generating authentic dopaminergic neurons in defined media from human PSC, and have determined the time point at which dopaminergic neurons can be frozen, shipped, thawed and transplanted without compromising their ability to mature and provide therapeutic benefit in animal models. Our process has been successfully transferred to a GMP facility and we have manufactured multiple lots of GMP-equivalent cells using this process. Importantly, we have shown functional equivalency of the manufactured cells in appropriate models. The tools, reagents and processes we have developed in this project allow us to move towards translational therapy and establish processes that could be applied to future IND-enabling projects. In addition, the processes we have developed would be of benefit to the CIRM community.
  • CIRM Progress Report Part A: Scientific Progress
  • I. Project Overview
  • During the past three years (36 months) we have successfully completed the milestones defined in the NGA for this grant. In brief, we have selected 1 clinically compliant ESC line H14 (and a back-up line H9), which have shown reproducible, efficient differentiation to dopaminergic neurons at lab scale. We have performed in vitro and in vivo characterization as defined in the NGA and guided by our discussion with our program officer at CIRM. We have determined the time point at which dopaminergic precursors (14 days after the NSC stage) can be frozen, shipped, thawed and transplanted without compromising their ability to mature and provide therapeutic benefit in animal models. Importantly, we have evaluated efficacy of cryopreserved dopaminergic precursors manufactured by the GMP-compatible process in a rodent PD model and shown functional recovery up to 6 months post transplantation as well as survival of dopaminergic neurons.
  • In the meanwhile we have successfully transferred the process of generating transplant ready dopaminergic neurons to the manufacture facilities at City of Hope (COH). They have adapted and optimized our protocols and have established GMP-compatible protocols for the culture of ESC-NSC and for differentiating NSC to Stage 3, Day 14 DA precursors for transplantation. During this reporting period (36 month), we have tested the equivalency of these lots and confirmed that lots manufactured at COH are consistent and similar to cells produced in the laboratory.
  • Our effort resulted in two important manuscripts in Cytotherapy:
  • 1. Liu, Q., Pedersen, OZ., Peng, J., Couture, LA., Rao, MS., and Zeng, X. Optimizing dopaminergic differentiation of pluripotent stem cells for the manufacture of dopaminergic neurons for transplantation. Cytotherapy. 2013 Aug;15(8):999-1010.
  • 2. Peng, J., Liu, Q., Rao, MS., and Zeng, X. Survival and engraftment of dopaminergic neurons manufactured by a GMP-compatible process. Cytotherapy. 2014 Sep;16(9):1305-12.

A hESc-based Development Candidate for Huntington's Disease

Funding Type: 
Early Translational II
Grant Number: 
TR2-01841
ICOC Funds Committed: 
$3 799 817
Disease Focus: 
Huntington's Disease
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 
Huntington’s disease (HD) is a devastating degenerative brain disease with a 1 in 10,000 prevalence that inevitably leads to death. These numbers do not fully reflect the large societal and familial cost of HD, which requires extensive caregiving. HD has no effective treatment or cure and symptoms unstoppably progress for 15-20 years, with onset typically striking in midlife. Because HD is genetically dominant, the disease has a 50% chance of being inherited by the children of patients. Symptoms of the disease include uncontrolled movements, difficulties in carrying out daily tasks or continuing employment, and severe psychiatric manifestations including depression. Current treatments only address some symptoms and do not change the course of the disease, therefore a completely unmet medical need exists. Human embryonic stem cells (hESCs) offer a possible long-term treatment approach that could relieve the tremendous suffering experienced by patients and their families. HD is the 3rd most prevalent neurodegenerative disease, but because it is entirely genetic and the mutation known, a diagnosis can be made with certainty and clinical applications of hESCs may provide insights into treating brain diseases that are not caused by a single, known mutation. Trials in mice where protective factors were directly delivered to the brains of HD mice have been effective, suggesting that delivery of these factors by hESCs may help patients. Transplantation of fetal brain tissue in HD patients suggests that replacing neurons that are lost may also be effective. The ability to differentiate hESCs into neuronal populations offers a powerful and sustainable alternative for cell replacement. Further, hESCs offer an opportunity to create cell models in which to identify earlier markers of disease onset and progression and for drug development. We have assembled a multidisciplinary team of investigators and consultants who will integrate basic and translational research with the goal of generating a lead developmental candidate having disease modifying activity with sufficient promise to initiate IND-enabling activities for HD clinical trials. The collaborative research team is comprised of investigators from multiple California institutions and has been assembled to maximize leverage of existing resources and expertise within the HD and stem cell fields.
Statement of Benefit to California: 
The disability and loss of earning power and personal freedom resulting from Huntington's disease (HD) is devastating and creates a financial burden for California. Individuals are struck in the prime of life, at a point when they are their most productive and have their highest earning potential. As the disease progresses, individuals require institutional care at great financial cost. Therapies using human embryonic stem cells (hESCs) have the potential to change the lives of hundreds of individuals and their families, which brings the human cost into the thousands. For the potential of hESCs in HD to be realized, a very forward-thinking team effort will allow highly experienced investigators in HD, stem cell research and clinical trials to come together and identify a lead development candidate for treatment of HD. This early translation grant will allow for a comprehensive and systematic evaluation of hESC-derived cell lines to identify a candidate and develop a candidate line into a viable treatment option. HD is the 3rd most prevalent neurodegenerative disease, but because it is entirely genetic and the mutation known, a diagnosis can be made with certainty and clinical applications of hESCs may provide insights into treating brain diseases that are not caused by a single, known mutation. We have assembled a strong team of California-based investigators to carry out the proposed studies. Anticipated benefits to the citizens of California include: 1) development of new human stem cell-based treatments for HD with application to other neurodegenerative diseases such as Alzheimer's and Parkinson's diseases that affect thousands of individuals in California; 2) improved methods for following the course of the disease in order to treat HD as early as possible before symptoms are manifest; 3) transfer of new technologies and intellectual property to the public realm with resulting IP revenues coming into the state with possible creation of new biotechnology spin-off companies; and 4) reductions in extensive care-giving and medical costs. It is anticipated that the return to the State in terms of revenue, health benefits for its Citizens and job creation will be significant.
Progress Report: 
  • Huntington’s disease (HD) is a devastating degenerative brain disease with a 1 in 10,000 prevalence that inevitably leads to death. Because HD is genetically dominant, the disease has a 50% chance of being inherited by the children of patients. Symptoms of the disease include uncontrolled movements, difficulties in carrying out daily tasks or continuing employment, and severe psychiatric manifestations including depression. Current treatments only address some symptoms and do not change the course of the disease, therefore a completely unmet medical need exists. Human embryonic stem cells (hESCs) offer a possible long-term treatment approach that could relieve the tremendous suffering experienced by patients and their families. Because HD is entirely genetic and the mutation known, a diagnosis can be made with certainty and clinical applications of hESCs may provide insights into treating brain diseases that are not caused by a single, known mutation. The ability to differentiate hESCs into neuronal populations offers a powerful and sustainable treatment opportunity. We have established the multidisciplinary team of investigators and consultants to integrate basic and translational research with the goal of generating a lead developmental candidate having disease modifying activity with sufficient promise to initiate IND-enabling activities for HD clinical trials.
  • In preliminary experiments, the transplantation of mouse neural stem cells, which survived in the brain for the four week period of the trial, provided protective effects in delaying disease progression in an HD mouse model and increased production of protective molecules in the brains of these mice. In the first year, the team has developed and established methods to differentiate hESCs into neural, neuronal and astrocyte precursors to be used for transplantation and has determined the correct cells to use that can be developed for future clinical development of these cells. In initial studies during this year, transplantation of neural stem cells (NSCs) provided both neurological and behavioral benefit to a HD mouse model. In addition, neuroprotective molecules were increased. Three immunosuppression regimens were tested to optimize methods for next stage preclinical trials. Finally, breeding of the three different HD mouse models has been initiated. Taken as a whole, progress supports the feasibility of the CIRM-funded studies to transplant differentiated hESCs into HD mice for preclinical development with the ultimate goal of initiating IND-enabling activities for HD clinical trials.
  • Huntington’s disease (HD) is a devastating degenerative brain disease with a 1 in 10,000 prevalence that inevitably leads to death. Because HD is genetically dominant, the disease has a 50% chance of being inherited by the children of patients. Symptoms of the disease include uncontrolled movements, difficulties in carrying out daily tasks or continuing employment, and severe psychiatric manifestations including depression. Current treatments only address some symptoms and do not change the course of the disease, therefore a completely unmet medical need exists. Human embryonic stem cells (hESCs) offer a possible long-term treatment approach that could relieve the tremendous suffering experienced by patients and their families. Because HD is entirely genetic and the mutation known, a diagnosis can be made with certainty and clinical applications of hESCs may provide insights into treating brain diseases that are not caused by a single, known mutation. The ability to differentiate hESCs into neuronal populations offers a powerful and sustainable treatment opportunity. We have established the multidisciplinary team of investigators and consultants to integrate basic and translational research with the goal of generating a lead developmental candidate having disease modifying activity with sufficient promise to initiate IND-enabling activities for HD clinical trials.
  • We previously performed transplantation of human neural stem cells into an HD mouse model and found that a subset of cells survived in the brain for the four week period of the trial, providing protective effects in delaying disease progression. In the past year, we have increased production and characterization of human neural stem cells (hNSCs) into neuronal (hNPC) and astrocyte (hAPC) precursors to be used for transplantation and optimized methods for shipping and implantation. Immunosuppression regimens were improved to optimize cell survival of implanted cells in HD mice. Transplantation of both human NSCs and NPCs are neuroprotective to HD mice and transplantation of hAPCs is in progress. Once completed, the cell giving the greatest protective benefit will be transplanted into mice that display slower progression over a longer time frame to validate and optimize approach for subsequent human application. All three HD mouse models have been bred and are ready for stem cell transplants. Taken as a whole, progress supports the feasibility of the CIRM-funded studies to transplant differentiated hESC-derived cell types into HD mice for preclinical development with the ultimate goal of identifying a lead candidate cell type and initiating IND-enabling activities for HD clinical trials.
  • Huntington’s disease (HD) is a devastating degenerative brain disease with a 1 in 10,000 prevalence that inevitably leads to death. Because HD is genetically dominant, the disease has a 50% chance of being inherited by the children of patients. Symptoms of the disease include uncontrolled movements, difficulties in carrying out daily tasks or continuing employment, and severe psychiatric manifestations including depression. Current treatments only address some symptoms and do not change the course of the disease, therefore a completely unmet medical need exists. Human embryonic stem cells (hESCs) offer a possible long-term treatment approach that could relieve the tremendous suffering experienced by patients and their families. Because HD is entirely genetic and the mutation known, a diagnosis can be made with certainty and clinical applications of hESCs may provide insights into treating brain diseases that are not caused by a single, known mutation. The ability to differentiate hESCs into neuronal populations offers a powerful and sustainable treatment opportunity. We have established the multidisciplinary team of investigators and consultants to integrate basic and translational research with the goal of generating a lead developmental candidate having disease modifying activity with sufficient promise to initiate Investigational New Drug (IND) enabling activities for HD clinical trials.
  • We have completed several rounds of transplantation of human neural stem cells into an HD mouse model and found that the cells survived in the brain for the four-week period of the trial, provided protective effects in delaying disease progression and increased production of protective molecules in the brains of these mice. In the last year the team differentiated hESCs into neural, neuronal and astrocyte precursors and performed transplantation studies to determine the best cell candidate to use and develop for future clinical work. We determined that the human neural stem cells produce the most robust effect. We have now selected a GMP grade hNSC line that will be carried forward for further testing in both rapidly progressing and slower progressing HD mice, as well as in mouse preclinical dosing studies. Taken as a whole, progress supports the feasibility of the CIRM-funded studies to transplant differentiated hESCs into HD mice for preclinical development with the ultimate goal on initiating IND-enabling activities for HD clinical trials.

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