Neurological Disorders

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

Stem Cell Mechanisms Governing Discrete Waves of Gliogenesis in the Childhood Brain

Funding Type: 
Basic Biology IV
Grant Number: 
RB4-06093
ICOC Funds Committed: 
$1 264 248
Disease Focus: 
Neurological Disorders
Pediatrics
Stem Cell Use: 
Adult Stem Cell
oldStatus: 
Active
Public Abstract: 
White matter is the infrastructure of the brain, providing conduits for communication between neural regions. White matter continues to mature from birth until early adulthood, particularly in regions of brain critical for higher cognitive functions. However, the precise timing of white matter maturation in the various neural circuits is not well described, and the mechanisms controlling white matter developmental/maturation are poorly understood. White matter is conceptually like wires and their insulating sheath is a substance called myelin. It is clear that neural stem and precursor cells contribute significantly to white matter maturation by forming the cells that generate myelin. In the proposed experiments, we will map the precise timing of myelination in the human brain and changes in the populations of neural precursor cells that generate myelin from birth to adulthood and define mechanisms that govern the process of white matter maturation. The resulting findings about how white matter develops may provide insights for white matter regeneration to aid in therapy for diseases such as cerebral palsy, multiple sclerosis and chemotherapy-induced cognitive dysfunction.
Statement of Benefit to California: 
Diseases of white matter account for significant neurological morbidity in both children and adults in California. Understanding the cellular and molecular mechanisms that govern white matter development the may unlock clues to the regenerative potential of endogeneous stem and precursor cells in the childhood and adult brain. Although the brain continues robust white matter development throughout childhood, adolescence and young adulthood, relatively little is known about the mechanisms that orchestrate proliferation, differentiation and functional maturation of neural stem and precursor cells to generate myelin-forming oligodendrocytes during postnatal brain development. In the present proposal, we will define how white matter precursor cell populations vary with age throughout the brain and determine if and how neuronal activity instructs neural stem and precursor cell contributions to human white matter myelin maturation. Disruption of white matter myelination is implicated in a range of neurological diseases, including cerebral palsy, multiple sclerosis, cognitive dysfunction from chemotherapy exposure, attention deficit and hyperactivity disorder (ADHD) and even psychiatric diseases such as schizophrenia. The results of these studies have the potential to elucidate clues to white matter regeneration that may benefit hundreds of thousands of Californians.
Progress Report: 
  • Formation of the insulated fiber infrastructure of the human brain (called "myelin") depends upon the function of a precursor cell type called "oligodendrocyte precursor cells (OPC)". The first Aim of this study seeks to determine how OPCs differ from each other in different regions of the brain, and over different ages. Understanding this heterogeneity is important as we explore the regenerative capacity of this class of precursor cells. We have, in the past year, isolated OPCs from various regions of the human brain from individuals at various ages and are studying the molecular characteristics of these precursor cells at the single cell level in order to define distinct OPC subpopulations. We have also begun to study the functional capabilities of OPCs isolated from different brain regions. The second Aim of this study seeks to understand how interactions between electrically active neurons and OPCs affect OPC function and myelin formation. We have found that when mouse motor cortex neurons "fire" signals in such a way as to elicit a complex motor behavior, much as would happen when one practices a motor task, OPCs within that circuit respond and myelination increases. This affects the function of that circuit in a lasting way. These results indicate that neurons and OPCs interact in important ways to modulate myelination and supports the hypothesis that OPC function may play a role in learning.
  • Sending neural impulses quickly down a long nerve fiber requires a specialized type of insulation called myelin, made by a cell called an oligodendrocyte that wraps itself around neuronal projections. Myelin-insulated nerve fibers make up the “white matter” of the brain, the vast tracts that connect one information-processing area of the brain to another. We have now shown that neuronal activity prompts oligodendrocyte precursor cell (OPC) proliferation and differentiation into myelin-forming oligodendrocytes. Neuronal activity also causes an increase in the thickness of the myelin sheaths within the active neural circuit, making signal transmission along the neural fiber more efficient. This was found to be true in both juvenile and in adult brains Metaphorically, it’s much like a system for improving traffic flow along roadways that are heavily used. And as with a transportation system, improving the routes that are most productive makes the whole system more efficient.
  • Interestingly, some parts of the neural circuit studied showed evidence of myelin-forming precursor cell response to neuronal activity, while other parts of the active circuit did not. In related work, we are making progress towards understanding how OPCs differ in various regions of the brain, examining the molecular heterogeneity of human OPCs at a single cell level.

Functional Neural Relay Formation by Human Neural Stem Cell Grafting in Spinal Cord Injury

Funding Type: 
Early Translational III
Grant Number: 
TR3-05628
ICOC Funds Committed: 
$4 699 569
Disease Focus: 
Spinal Cord Injury
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 
We aim to develop a novel stem cell treatment for spinal cord injury (SCI) that is substantially more potent than previous stem cell treatments. By combining grafts of neural stem cells with scaffolds placed in injury sites, we have been able to optimize graft survival and filling of the injury site. Grafted cells extend long distance connections with the injured spinal cord above and below the lesion, while the host spinal cord also sends inputs to the neural stem cell implants. As a result, new functional relays are formed across the lesion site. These result in substantially greater functional improvement than previously reported in animal studies of stem cell treatment. Work proposed in this grant will identify the optimal human neural stem cells for preclinical development. Furthermore, in an unprecedented step in spinal cord injury research, we will test this treatment in appropriate preclinical models of SCI to provide the greatest degree of validation for human translation. Successful findings could lead to clinical trials of the most potent neural stem cell approach to date.
Statement of Benefit to California: 
Spinal cord injury (SCI) affects approximately 1.2 million people in the United States, and there are more than 11,000 new injuries per year. A large number of spinal cord injured individuals live in California, generating annual State costs in the billions of dollars. This research will examine a novel stem cell treatment for SCI that could result in functional improvement, greater independence and improved life styles for injured individuals. Results of animal testing of this approach to date demonstrate far greater functional benefits than previous stem cell therapies. We will generate neural stem cells from GMP-compatible human embryonic stem cells, then test them in the most clinically relevant animal models of SCI. These studies will be performed as a multi-center collaborative effort with several academic institutions throughout California. In addition, we will leverage expertise and resources currently in use for another CIRM-funded project for ALS, thereby conserving State resources. If successful, these studies will form the basis for clinical trials in a disease of great unmet medical need, spinal cord injury. Moreover, the development of this therapy would reduce costs for clinical care while bringing novel biomedical resources to the State.
Progress Report: 
  • In the first 12 months of this project we have made important progress in the following areas:
  • 1) Identified the lead embryonic stem cell type for potential use in a translational clinical program.
  • 2) Replicated the finding that implants of ES-derived neural progenitor cells from this lead cell type extend axons out from the spinal cord lesion site in very high numbers and over very long distances.
  • 3) Begun efforts to scale this work to larger animal models of spinal cord injury.
  • Very good progress has been made in the last year on this project. We are attempting to address a great unmet medical need to develop effective therapies for human spinal cord injury (SCI). We aim to develop and optimize a pluripotent neural stem cell line for grafting to sites of spinal cord injury, and develop this line for clinical translation. Unlike other programs of stem cell therapy for SCI, we are transplanting neural stem cells directly into the injury site, in high numbers, and we observe very extensive growth of axons both into and out of the graft. The amount of axon growth in this model is substantially greater than that observed with other approaches to the injured spinal cord, including approaches currently in clinical trials. Accordingly, we believe that our approach provides a substantially greater opportunity to improve outcomes after SCI.
  • In the last year, we have identified a lead stem cell line for potential human translation, and validated its ability to engraft to the injured spinal cord. We have observed that human neural stem cells, grafted into mice and rats, exhibit a human time frame for maturation and growth: cells require at least one year to develop and mature. This knowledge is very important for planning human clinical trials.
  • Remaining work will characterize the long term safety and efficacy of these cells in rodent and large animal models of SCI.

Understanding the role of LRRK2 in iPSC cell models of Parkinson's Disease

Funding Type: 
Basic Biology III
Grant Number: 
RB3-02221
ICOC Funds Committed: 
$1 482 822
Disease Focus: 
Parkinson's Disease
Neurological Disorders
Stem Cell Use: 
iPS Cell
oldStatus: 
Active
Public Abstract: 
The goal of this research is to utilize novel research tools to investigate the molecular mechanisms that cause Parkinson’s disease (PD). The proposed work builds on previous funding from CIRM that directed the developed patient derived models of PD. The majority of PD patients suffer from sporadic disease with no clear etiology. However some PD patients harbor specific inherited mutations have been shown to cause PD. The most frequently observed form of genetic parkinsonism is caused by the LRRK2 G2019S mutation it the most common. This mutation accounts for approximately 1.5-2% of patients with apparently sporadic PD, increasing to 4-6% of patients with a family history of PD, and even higher in isolated populations. Importantly, LRRK2 induced PD is clinically and pathologically largely indistinguishable from sporadic PD. This proposal focuses on studying the most frequent cause of familial PD and induces disease that is clinically and pathologically identical to sporadic PD cases. It is likely that LRRK2 regulates a pathway(s) that is important in the more common sporadic form of PD as well. Therefore by employing relevant models of PD, we hope to drive the biological understanding of LRRK2 in a direction that facilitates the development of disease therapeutics in the future. We ascertained patients harboring mutations in LRRK2 [heterozygous (+/G2019S) and homozygous (G2019S/G2019S)] as well as sporadic cases and age matched controls. We have successfully derived iPSCs from each genotype and differentiated these to DA neurons. We will use these as a model system to investigate these LRRK2 based models of PD. We will adapt current biochemical assays of LRRK2, which are source material intensive, to the small culture volumes required for the differentiation of iPSCs to DA neurons. This is a crucial necessity for development for utilizing iPSC derived DA neurons as tractable models of LRRK2 based PD. We will then probe the roles of LRRK2 in neuronal cell differentiation and survival. We will also ask whether the mutant LRRK2 induces changes in autophagy, as this has been postulated as a mechanism of LRRK2 induced pathogenesis. By studying wild-type and disease mutant LRRK2, in DA models of PD we hope to provide crucial understanding of the role mutant LRRK2 has in disease.
Statement of Benefit to California: 
It is estimated that by the year 2030, 75,000-120,000 Californians will be affected by Parkinson’s disease. Currently, there is no cure, early detection mechanism, preventative treatment, or effective way to slow disease progression. The increasing disability caused by the progression of disease burdens the patients, their caregivers as well as society in terms of healthcare costs. The majority of PD patients suffer from sporadic disease with no clear etiology, and a in a handful of these patients specific inherited mutations have been shown to cause PD. The most frequently mutated gene is called Leucine Rich Repeat Kinase 2 (LRRK2). Our goal is to study the mutated gene product in patient based models of Parkinson’s disease. In previous CIRM funding, we have developed patient derived induced pluripotent stem cells (iPSCs) from patients harboring mutations in LRRK2. We have been successful in differentiating populations these iPSCs into the neurons that are depleted in PD. The next step is to utilize these cells as models of mutation induced PD ‘in a dish’. We will employ these pertinent disease models to answer basic biology questions that remain about the function of LRRK2. This project brings together scientists previously funded by CIRM with scientists well versed in the study of LRRK2. This multidisciplinary approach to studying the causes of PD is a natural benefit to the State of California and its citizens. By bringing a better understanding of the role of LRRK2 in the cells that are lost in the progression of PD, we will bring more concrete knowledge of PD as a whole, bringing more hope for the development of a therapeutic for disease.
Progress Report: 
  • The overarching goal of this work is to utilize models of Parkinson's disease (PD) that originate from cells of PD affected patients harboring mutations within the LRRK2 gene so that we may discern the role of mutated LRRK2 in disease. Mutations in LRRK2 are the most common cause of familial PD. The disease presentation of patients with LRRK2 mutation is typically clinically indistinguishable from sporadic PD cases, making the onset of disease due to LRRK2 dysfunction clinically relevant. We have employed stem cells derived from these patients to generate neuronal cells in which we can determine the roles of LRRK2 in the PD mutated and the unmutated state. We have focused on a cellular process called autophagy that regulates the cell response to nutrient deprivation and plays a role in the selective degradation of proteins within the cell.
  • In the first year of funding we have analyzed the expression of the protein LRRK2 in induced pluripotent stem cells, neuronal precursor cells and have begun to differentiate the neuronal precursors to dopaminergic cells of the type lost in PD (a difficult task in itself). We have applied a novel method for detection of LRRK2 in situ by marrying the protein detection of antibodies and the sensitivity of nucleic acid amplification. We will continue to develop this methodology for maximum sensitivity to LRRK2. We have established assays to assess the effects of the LRRK2 mutant on autophagy that are relevant to PD and neurological diseases in general. We have met or made great progress on most of our anticipated milestones and are eager to proceed to the next phase of the project.
  • The overarching goal of this work is to utilize stem cell based models of Parkinson's disease (PD) derived from cells of PD affected patients that harbor mutations in the LRRK2 gene so that we may elucidate the deleterious role of mutated LRRK2 in disease. Mutations in LRRK2 are the most common cause of familial PD. The disease presentation for these patients with LRRK2 mutation is typically clinically similar to those with sporadic disease, making the onset of disease due to LRRK2 dysfunction clinically relevant. We have utilized stem cells harboring a mutation in LRRK2 and also daughter cells of that line in which genomic editing techniques have been applied to correct the PD mutation or disrupt the LRRK2 gene. We have generated the same kind of cells in culture that are lost during PD and hope that next, we can determine how these mutations that eventually cause disease disrupt normal neuronal function. We have made great progress in the understanding the expression of LRRK2 in early differentiation of stem cells to neurons and his will inform our future studies on mutation caused dysfunctions.
  • The overarching goal of this work is to utilize stem cell based models of Parkinson's disease (PD) derived from cells of PD affected patients that harbor mutations in the LRRK2 gene so that we may elucidate the deleterious role of mutated LRRK2 in disease. Mutations in LRRK2 are the most common cause of familial PD. The disease presentation for these patients with LRRK2 mutation is typically clinically similar to those with sporadic disease, making the onset of disease due to LRRK2 dysfunction clinically relevant. We have utilized stem cells harboring a mutation in LRRK2 and also daughter cells of that line in which genomic editing techniques have been applied to correct the PD mutation or disrupt the LRRK2 gene. We have generated the same kind of cells in culture that are lost during PD and hope that next, we can determine how these mutations that eventually cause disease disrupt normal neuronal function. We have made great progress in the understanding the expression of LRRK2 in early differentiation of stem cells to neurons and this will inform our future studies on mutation caused dysfunctions.
  • During our project we achieved several scientific goals. Our project was to utilize Parkinson’s disease patient derived stem cells to model their disease. The particular cells we used were derived from patients with mutations in the LRRK2 gene, which is the most common cause of inherited Parkinson’s disease. At the end of our funding we can report proficiency in induced pluripotent stem cell (iPSC) differentiation to dopaminergic cells protocols in a workflow that allows higher throughput analysis. Dopaminergic cells are types of cells lost in the brain in Parkinson’s disease and if we study cells from patients with this mutation it will likely yield insight into the processes of disease onset and progression. This funding enabled collaboration with the laboratory of Dr. Schuele here at the Parkinson’s Institute to use cells generated in other CIRM funding (ETI-0246 and RT2-019665). The cell lines we used were edited to “fix” the genetic mutation in the LRRK2 gene and also to delete (or knockout) the LRRK2 gene. We used these cells and determined that loss of LRRK2 imparts an increased propensity for dopaminergic differentiation potential for knockouts over mutant and wild-type cells. Also, we determined that LRRK2 inhibitors do not negatively impact the generation of dopaminergic cells in cell culture modeling. We will continue to unravel the roles of LRRK2 in Parkinson’s disease using these pertinent, patient centric models.

Development of small molecule screens for autism using patient-derived iPS cells

Funding Type: 
Tools and Technologies II
Grant Number: 
RT2-01906
ICOC Funds Committed: 
$1 884 808
Disease Focus: 
Autism
Neurological Disorders
Pediatrics
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 
Autism Spectrum Disorders (ASDs) are a heritable group of neuro-developmental disorders characterized by language impairments, difficulties in social integrations, and the presence of stereotyped and repetitive behaviors. There are no treatments for ASDs, and very few targets for drug development. Recent evidence suggests that some types of ASDs are caused by defects in calcium signaling during development of the nervous system. We have identified cellular defects in neurons derived from induced pluripotent stem cells (iPSCs) from patients with Timothy Syndrome (TS), caused by a rare mutation in a calcium channel that leads to autism. We propose to use cells carrying this mutant calcium channel to identify drugs that act on calcium signaling pathways that are involved in ASDs. Our research project has three aims. First, we will determine whether known channel modulators reverse the cellular defects we observe in cells from TS patients. It is possible that we will find that existing drugs already approved for use in humans might be effective for treating this rare but devastating disorder. Our second aim is to determine whether screens using neuronal cells derived from ASD patients can be used to identify calcium signaling modulators. A bottleneck to therapy development for ASDs has been the lack of appropriate in vitro models for these disorders, and we would like to determine whether our studies could serve as the basis for a new type of screen in human neurons. Our third aim is to identify signaling molecules that might be affected in patients with ASDs, which could be targets for future drug discovery. There is increasing evidence that several types of ASDs are caused by defects in neuronal activity and calcium signaling. More specifically, the CaV1.2 calcium channel that we are studying has been implicated in syndromic and non-syndromic forms of autism, and also in schizophrenia and bipolar disorder. One of the more exciting aspects of our screen of neurons with a mutation in CaV1.2 is that it gives us a tool to explore calcium-mediated signaling pathways that are defective in ASDs. We will try to modify calcium signaling in neurons from ASD patients by changing the expression of proteins that are known to affect calcium signaling in other contexts. These experiments will identify targets that are active in human neurons and that affect cellular phenotypes that are defective in ASD. In summary, the work described in this proposal constitutes a critical step to fulfilling the promise that reprogramming of patient-specific cells offers for the treatment of neuropsychiatric disorders such as autism. Our studies will identify lead compounds that could be tested in the clinic for a rare form of autism, and novel molecular targets for therapeutic development in the future. Importantly, these studies will provide a proof of principle that iPSC-derived cells are valuable for drug discovery for neuropsychiatric disorders.
Statement of Benefit to California: 
Autism Spectrum Disorders (ASDs) affect approximately 1 in 110 children in California. In addition to the devastating effects that ASDs have on the families of affected individuals, treating and educating people with ASDs imposes a heavy economic burden on the state. In 2007, almost 35,000 individuals with autism were receiving services from the California Regional Centers, and the number was expected to rise to 50,000 by last year. Recent estimates suggest that the lifetime cost of caring for an individual with an ASD can exceed $3 million. In spite of their impact on our society, there are currently no effective therapies for ASDs. Our lack of cellular and molecular tools to study these disorders means that there are no good targets for drug screening, so there are very limited prospects for developing effective pharmacological treatments in the near future. New drug discovery paradigms are needed to help develop therapies for these neuropsychiatric conditions. The research described in this proposal could have a dramatic impact on drug discovery methods for ASDs. First, we hope to identify drugs that are effective in treating Timothy Syndrome, a rare form of autism caused by an electrophysiological defect in a calcium channel. Second, we aim to develop new tools to explore calcium-mediated signaling pathways that are defective in ASDs. If successful, our research will identify a family of molecular targets that will be useful for developing therapies for ASDs in the future.
Progress Report: 
  • Autism Spectrum Disorders (ASDs) are a heritable group of neuro-developmental disorders characterized by language impairments, difficulties in social integrations, and the presence of stereotyped and repetitive behaviors. There are no treatments for ASDs, and very few targets for drug development. The goal of this CIRM project is to develop a series of in vitro screens for drugs that might affect the underlying cellular defects in ASDs.
  • Since ASDs are uniquely human, we proposed to design, optimize and conduct high-throughput chemical screens using human neurons derived from induced pluripotent stem cells (iPSCs). Our lab identified cellular defects in neurons derived from patients with Timothy Syndrome (TS), a syndromic disorder often presenting with autism that is caused by a rare mutation in a calcium channel. In our project, we proposed to develop in vitro screening assays for ASDs based on these TS phenotypes, and to screen these assays to identify drugs that might affect behavioral symptoms of autism. In the first year of this award, we conducted preliminary screens and found that certain calcium channel modulators reverse some of the differentiation defects that we observe in these cells. We also extended observations that we had made in mice and showed that TS neurons have defects in the structure and length of their dendrites, measurable features that we can use as the basis for additional drug screens. We have therefore progressed within the aims of the original award.
  • For the remainder of the grant, however, we are proposing to broaden the scope of this project to include iPSC-based screens using neurons from patients with more prevalent forms of ASDs. In other research in our lab, we have characterized phenotypes in neurons derived from patients with two other diseases that are more prevalent than TS: DiGeorge Syndrome (DGS) and Phelan-McDermid Syndrome (PMDS), two neurodevelopmental disorders resulting from deletions within chromosome 22 and patients present symptoms that often include autism. We have shown that these cells have defects in the length of their dendrites, in the structure and function of their synapses, and in their ability to transmit electrical impulses. We propose to broaden the scope of our work to develop screens for TS, DGS, and PMDS. These screens will serve as a basis for identifying drugs that lessen or reverse cellular defects in these disorders, and thus may lead to more generalized treatments for ASDs.
  • We believe that this research not only fulfills critical steps in the development of a novel test for potential ASD treatments, but demonstrates the power of iPSC technology for understanding the underlying mechanisms of neurological disorders. Expanding the scope of our original project will help us increase the impact of our studies on therapeutic development and on the understanding of the neurobiology of ASDs.
  • Autism Spectrum Disorders (ASDs) are a heritable group of neurodevelopmental disorders that affect the verbal, social, and behavioral abilities of affected individuals. There are no pharmacological treatments for ASDs, in part because of a lack of validated cellular and animal models for use in drug screens. The goal of this project is to develop and validate a cell-based high throughput screening method that we will use to identify therapies for ASDs.
  • Our laboratory has established methods for collecting skin samples from patients and reprogramming these cells into induced pluripotent stem (iPS) cells, which we then differentiate into neurons. We have characterized neurons from patients with ASDs, and identified cellular phenotypes that are amenable to high-throughput methods to identify drug targets. Our efforts in Year 2 of our CIRM funding have focused on Phelan-McDermid Syndrome (PMDS), an inherited progressive neurodevelopmental disorder characterized by developmental delay, absent or severely impaired speech, and an increased risk of autism. We have discovered that neurons from PMDS patients who have autism have defects in excitatory synaptic transmission caused by the loss of one copy of the gene Shank3. Shank3 lies in the region of Chromosome 22 that is deleted in PMDS, and is important for the development of synapses. Based on our studies, PMDS neurons can be distinguished from their wildtype counterparts by low expression levels of Shank3 measured by quantitative PCR, decreased number of excitatory synapses labeled by immunocytochemistry and imaged with a microscope, and reduced excitatory cellular currents measured electrophysiologically. Each of these phenotypes is amenable to high throughput screening of therapeutic compounds. We tested several candidate therapeutics and found that prolonged treatment with the growth factor IGF-1 partially reverses the defects we have discovered in PMDS neurons. While IGF-1 is highly bioactive and therefore not an ideal drug candidate, it can be used to validate our screening method.
  • We are currently running trials to select the best phenotype and assay for larger-scale screening. In parallel, we have developed protocols to culture large numbers of iPSC-derived neurons for high throughput screens, and we are growing and banking working stocks of PMDS and control neurons. These experiments will help us identify drug candidates for PMDS, and will represent a significant advance in HTS approaches for the testing of ASD therapies using iPSC-based systems.
  • Autism Spectrum Disorders (ASDs) are a heritable group of neurodevelopmental disorders that affect the verbal, social, and behavioral ability of affected in individual. There are no treatments for ASD, in part because the biological basis for the disorders are not know. In addition, there are no methods for screening drugs that may be therapeutic. The goal of this project was to develop screening assays based on stem cells that were derived from individuals with autism.
  • Using skin samples from affected individuals, our laboratory was able to generate induced pluripotent stem cells (iPSC) and use these stem cells to generate neurons. With CIRM support, we have now generated iPSC from many individuals, some of whom carry genetic alterations that cause autism. Work under this award focused on two genetic disorders, Timothy Syndrome (TS) and Phelan-McDermid Syndrome (PMDS). Both are inherited syndromes that affect several body systems and also greatly increase the risk of autism. In each case, we found that neurons from affected individuals displayed changes in the way neurons connect and communicate. The effects were pronounced in PMDS neurons, in part due to the loss of the Shank3 gene that is involved in the function of the excitatory synapse. Work in year 3 has focused on identifying a robust alteration in neuron function that can be used for drug screening.
  • One such phenotype was discovered and involves a change in the way calcium is utilized when neurons communicate by generating an electrical current. Using chemicals that detect calcium, fluorescent assays were developed that show a robust difference in calcium response in PMDS neurons relative to neurons from unaffected individuals. Adapting the fluorescent calcium reporter assay to a high-throughput format also required the invention of new stem cell culture methods for generating neurons that were more efficient and less costly. Ultimately, a novel strategy was developed that now permits the production of very large numbers of neurons that can be assayed in high throughput screens. A limited screen using candidate drugs has confirmed the utility of the assay and future work will utilize these assays in large scale screens for drugs that normalize or augment the synaptic defects.

Skin-derived precursor cells for the treatment of enteric neuromuscular dysfunction

Funding Type: 
Tools and Technologies III
Grant Number: 
RT3-07914
ICOC Funds Committed: 
$1 818 751
Disease Focus: 
Intestinal Disease
Pediatrics
Neurological Disorders
Stem Cell Use: 
Adult Stem Cell
Public Abstract: 
The intestine performs the essential function of absorbing food and water into the body. Without a functional intestine, children and adults cannot eat normal meals, and these patients depend on intravenous nutrition to sustain life. Many of these patients do not have a neural system that coordinates the function of the intestine. These patients have a poor quality of life, and the cost of medical care is over $200,000 per year for each patient. Stem cell therapies offer potential cures for these patients while avoiding the risks of invasive procedures and hazardous treatments. A novel approach to treat these patients is to use stem cells derived from the patient’s own skin to generate the neural system. This has been shown to be feasible in small animals, and the next step hinges on the demonstration of these results in a large animal model of intestinal dysfunction. We will develop a model in large animals that can be used to test the ability of skin-derived stem cells to form the neural system. Skin-derived stem cells will be isolated from large animal models and human skin to demonstrate their potential to generate a functional neural system. These cells will be transplanted into the animal model to determine the best way for these cells to make the intestine function properly. This research will gather critical information needed to begin a clinical trial using skin-derived cells to treat intestinal dysfunction.
Statement of Benefit to California: 
Gastrointestinal dysfunction destroys the lives of thousands of Californians. These Californians have frequent and prolonged hospitalizations and lost wages due to their chronic illness. It is estimated that the health care cost of Californians with gastrointestinal neuromuscular dysfunction is over 400 million dollars annually. Currently, most of these patients are covered by the state’s insurance agency. Stem cell therapies offer potential cures for these patients and reduce this economic burden. The proposed research will obtain critical information needed to begin a clinical trial using skin-derived cells to treat patients with intestinal dysfunction. The California economy will significantly benefit from this research through the reduced costs for health care and increased quality of life of the affected Californians. Additionally, this work will add to the state’s growing stem cell industry and will increase employment opportunities and revenue by the state of California. The educational benefit to Californians involved in this research project will also maintain California’s position in leading the stem cell effort in the future.

Programming Human ESC-derived Neural Stem Cells with MEF2C for Transplantation in Stroke

Funding Type: 
Early Translational IV
Grant Number: 
TR4-06788
ICOC Funds Committed: 
$2 124 000
Disease Focus: 
Stroke
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 
The goal of this project is to produce a stem cell-based therapy for stroke (also known as an ischemic cerebral infarct). Stroke is the third leading cause of death in the USA, and a leading cause of disability among adults. Currently, there are no effective treatments once a stroke has occurred (termed completed stroke). In this proposal, we aim to develop human stem cells for therapeutic transplantation to treat stroke. Potential benefits will outweigh risks because only patients with severe strokes that have compromised activities of daily living to an extreme degree will initially be treated. Using a novel approach, we will generate stem cells that do not form tumors, but instead only make new nerve cells. We will give drugs to avoid rejection of the transplanted cells. Thus, the treatment should be safe. We will first test the cells in stroke models in rodents (mice and rats) in preparation for a human clinical trial. We will collect comprehensive data on the mice and rats to determine if the stem cells indeed become new nerve cells to replace the damaged tissue and to assess if the behavior of the mice and rats has improved. If successfully developed and commercialized, this approach has the potential for revolutionizing stroke therapy.
Statement of Benefit to California: 
The goal of this project is to produce a stem cell-based therapy for stroke (also known as an ischemic cerebral infarct). Stroke is the third leading cause of death in the State of California, and a leading cause of disability among adults. Currently, there are no effective treatments once a stroke has occurred (termed completed stroke), and the quality of life is severely compromised in those that survive the malady. In this proposal, we aim to develop human stem cells for therapeutic transplantation to treat stroke. Using a novel approach, we will generate stem cells that do not form tumors, but instead only make new nerve cells. If successfully developed and commercialized, this approach could provide a therapeutic candidate for the unmet medical need, which would have a tremendous impact on the quality of life for the patient, his or her family, and for the economic and emotional burden on the State of California and its citizens.
Progress Report: 
  • Stroke is the third leading cause of death in the USA and remains a great medical problem in California. Currently, there is no effective treatment for patients with a stroke who are seen several hours after the event. The goal of this project is to establish the feasibility of using a stem cell line for cell-replacement therapy to target stroke (cerebral ischemia). Using a novel, genetic pre-programming approach, we will generate human neural stem/progenitor cells (hNSC/NPCs) that are resistant to apoptotic cell death and destined to become nerve cells (or neurons). Our approach also avoids tumor formation, which can occur if stem cells that are not programmed to become neurons are injected into the brain. We will achieve our goals by introducing a constitutively active form of the transcription factor MEF2C (MEF2CA) into human embryonic stem cell (hESC)-derived hNPCs. For this purpose it is critical to identify a viral vector system that is the safest and most effective in producing MEF2CA-programmed hNPCs. We decided to use an adenoviral-associated virus (AAV) vector system because unlike other viral delivery methods (e.g., lentiviral, retroviral), an AAV system allows us to achieve MEF2CA expression in an integration-free and transient manner, as required for proper neuronal differentiation from NPCs. After in vitro characterization of these cells, including their neurogenic capacity, scalability etc., we will transplant them into rodent models of focal stroke. We are analyzing transplanted rats with immunohistochemical, electrophysiological, and behavioral methods to determine whether MEF2CA-programmed hNPCs can successfully differentiate into functional, integrated neurons into the host brain and ameliorate stroke-induced behavioral deficits. To assess the robustness of the AAV approach, we will also compare the results obtained from this system to those obtained using a hESC line that is stably programmed (resulting in permanent insertion of the MEF2C transgene into the genome of the cell, as opposed to transient MEF2C expression achieved with the AAV system). These studies will allow us to determine the effectiveness of the integration-free AAV system vs. stable integration of MEF2C on hNPCs developed for cell replacement therapy. During the current reporting period (Year 01), we have efficiently produced an AAV vector that transduces the MEF2CA transgene into hESC-derived NPCs. FACS analysis revealed that we have robustly infected the hNPCs with this AAV-based construct (~95% of cells infected). In vitro evaluation for protein and mRNA (through immunocytochemistry, and qRT-PCR assays) from the cells infected with AAV-MEF2CA revealed their neural progenitor cell identity and that the MEF2CA transgene is active in these cells. We have begun to transplant these cells into the brain of the spontaneously hypertensive (SHR) rat model of focal stroke. We have begun to compare the effects in stroke of the AAV-MEF2CA and stable-MEF2CA cell lines. In vitro characterization of the stable MEF2CA stem cell line demonstrated that we can differentiate these hNPCs into neurons. For example, protein, mRNA, and morphological analyses revealed robust differentiation of these hNPCs into mature cerebrocortical neurons. Electrophysiological analysis further confirmed the expression of functional neuronal channels and synaptic currents. Behavioral evaluation performed 12-weeks after transplant into the stroked brain with the stable-MEF2CA hNPC line vs. control revealed promising behavioral improvements in the MEF2CA-NPC transplanted group compared to control without apparent side effects.

Generation and characterization of high-quality, footprint-free human induced pluripotent stem cell lines from 3,000 donors to investigate multigenic diseases

Funding Type: 
hiPSC Derivation
Grant Number: 
ID1-06557
ICOC Funds Committed: 
$16 000 000
Disease Focus: 
Developmental Disorders
Genetic Disorder
Heart Disease
Infectious Disease
Alzheimer's Disease
Neurological Disorders
Autism
Respiratory Disorders
Vision Loss
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 
Induced pluripotent stem cells (iPSCs) have the potential to differentiate to nearly any cells of the body, thereby providing a new paradigm for studying normal and aberrant biological networks in nearly all stages of development. Donor-specific iPSCs and differentiated cells made from them can be used for basic and applied research, for developing better disease models, and for regenerative medicine involving novel cell therapies and tissue engineering platforms. When iPSCs are derived from a disease-carrying donor; the iPSC-derived differentiated cells may show the same disease phenotype as the donor, producing a very valuable cell type as a disease model. To facilitate wider access to large numbers of iPSCs in order to develop cures for polygenic diseases, we will use a an episomal reprogramming system to produce 3 well-characterized iPSC lines from each of 3,000 selected donors. These donors may express traits related to Alzheimer’s disease, autism spectrum disorders, autoimmune diseases, cardiovascular diseases, cerebral palsy, diabetes, or respiratory diseases. The footprint-free iPSCs will be derived from donor peripheral blood or skin biopsies. iPSCs made by this method have been thoroughly tested, routinely grown at large scale, and differentiated to produce cardiomyocytes, neurons, hepatocytes, and endothelial cells. The 9,000 iPSC lines developed in this proposal will be made widely available to stem cell researchers studying these often intractable diseases.
Statement of Benefit to California: 
Induced pluripotent stem cells (iPSCs) offer great promise to the large number of Californians suffering from often intractable polygenic diseases such as Alzheimer’s disease, autism spectrum disorders, autoimmune and cardiovascular diseases, diabetes, and respiratory disease. iPSCs can be generated from numerous adult tissues, including blood or skin, in 4–5 weeks and then differentiated to almost any desired terminal cell type. When iPSCs are derived from a disease-carrying donor, the iPSC-derived differentiated cells may show the same disease phenotype as the donor. In these cases, the cells will be useful for understanding disease biology and for screening drug candidates, and California researchers will benefit from access to a large, genetically diverse iPSC bank. The goal of this project is to reprogram 3,000 tissue samples from patients who have been diagnosed with various complex diseases and from healthy controls. These tissue samples will be used to generate fully characterized, high-quality iPSC lines that will be banked and made readily available to researchers for basic and clinical research. These efforts will ultimately lead to better medicines and/or cellular therapies to treat afflicted Californians. As iPSC research progresses to commercial development and clinical applications, more and more California patients will benefit and a substantial number of new jobs will be created in the state.
Progress Report: 
  • First year progress on grant ID1-06557, " Generation and Characterization of High-Quality, Footprint-Free Human Induced Pluripotent Stem Cell (iPSC) Lines From 3000 Donors to Investigate Multigenic Disease" has met all agreed-upon milestones. In particular, Cellular Dynamics International (CDI) has taken lease to approximately 5000 square feet of lab space at the Buck Institute for Research on Aging in Novato, CA. The majority of this space is located within the new CIRM-funded Stem Cell Research Building at the Buck Institute and was extensively reconfigured to meet the specific needs of this grant. All equipment, including tissue culture safety cabinets and incubators, liquid-handling robotics, and QC instrumentation have been installed and qualified. A total of 16 scientists have been hired and trained (13 in Production and 3 in Quality) and more than 20 Standard Operating Procedures (SOPs) have been developed and approved specifically for this project. These SOPs serve to govern the daily activities of the Production and Quality staff and help ensure consistency and quality throughout the iPSC derivation and characterization process. In addition, a Laboratory Information Management System (LIMS) had to be developed to handle the large amount of data generated by this project and to track all samples from start to finish. The first and most important phase of this LIMS project has been completed; additional functionalities will likely be added to the LIMS during the next year, but completion of phase 1 will allow us to enter full production mode on schedule in the first quarter of year 2. Procedures for the shipping, infectious disease testing, and processing of donor samples were successfully implemented with the seven Tissue Collectors. To date, over 700 samples have been received from these Tissue Collectors and derivation of the first 50 patient-derived iPSC lines has been completed on schedule. These cells have been banked in the Coriell BioRepository, also located at the Buck Institute. The first Distribution Banks will be available for commercial release during year 2.

Stem cell based small molecule therapy for Alzheimer's disease

Funding Type: 
Early Translational III
Grant Number: 
TR3-05669
ICOC Funds Committed: 
$1 673 757
Disease Focus: 
Alzheimer's Disease
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 
Over 6 million people in the US suffer from AD. There are no drugs that prevent the death of nerve cells in AD, nor has any drug been identified that can stimulate their replacement. Even if nerve cells could be replaced, the toxic environment of the brain will kill them unless they are protected by a drug. Therefore, drugs that stimulate the generation of new neurons (neurogenesis) alone will not be effective; a drug with both neurogenic and neuroprotective properties is required. With the ability to use cells derived from human embryonic stem cells (hESCs) as a screen for neurogenic compounds, it should now be possible to identify and tailor drugs for therapeutic use in AD. Our laboratory has developed a drug discovery scheme based upon using hESCs to screen drug candidates. We have recently identified a very potent drug that is exceptionally effective in rodent models of AD. However, this molecule needs to be optimized for human use. In this proposal, we will harness the power of hESCs to develop derivatives of J147 specifically tailored to stimulate neurogenesis and be neuroprotective in human cells. This work will optimize the chances for its true therapeutic potential in AD, and presents a unique opportunity to expand the use of hESCs for the development of a therapeutic for a disease for which there is no cure. This work could lead to a paradigm shift in the treatment of neurodegenerative disease.
Statement of Benefit to California: 
Over 6 million people in the US suffer from Alzheimer’s disease (AD). Unless a viable therapeutic is identified it is estimated that this number will increase to 16 million by 2050, with a cost of well over $1 trillion per year, overwhelming California and national health care systems. Among the top 10 causes of death, AD (6th) is the only one with no treatment available to prevent, cure or slow down the condition. An enormous additional burden to families is the emotional and physical stress of having to deal with a family member with a disease which is going to become much more frequent with our aging population. In this application we use new human stem cell technologies to develop an AD drug candidate based upon a strong lead compound that we have already made that stimulates the multiplication of nerve precursor cells derived from human embryonic stem cells. This approach presents a unique opportunity to expand the use of human embryonic stem cells for the development of a therapeutic for a disease for which there is no cure, and could lead to a paradigm shift in the treatment of neurodegenerative disease. Since our AD drug discovery approach is fundamentally different from the unsuccessful approaches used by the pharmaceutical industry, it could also stimulate new biotech. The work in this proposal addresses one of the most important medical problems of California as well as the rest of the world, and if successful would benefit all.
Progress Report: 
  • Introduction: Over 6 million people in the US suffer from AD. There are no drugs that prevent the death of nerve cells in AD, nor has any drug been identified that can stimulate their replacement. Even if nerve cells could be replaced, the toxic environment of the brain will kill them unless they are protected by a drug. Therefore, drugs that stimulate the generation of new neurons (neurogenesis) alone will not be effective; a drug with both neurogenic and neuroprotective properties is required. With the ability to use cells derived from human embryonic stem cells (hESCs) as a screen for neurogenic compounds, it should now be possible to identify and tailor drugs for therapeutic use in AD. This is the overall goal of this application.
  • Year One Progress: Using a novel drug discovery paradigm, we have made a very potent drug called J147 that is exceptionally effective in rodent models of AD and also stimulates neurogenesis in both young and very old mice. Very few, if any, drugs or drug candidates are both neuroprotective and neurogenic, particularly in old animals. In the first year of this application we harnessed the power of hESCs and medicinal chemistry to develop derivatives of J147 specifically tailored to stimulate neurogenesis and be neuroprotective in human cells. Using iterative chemistry, we synthesized over 200 new compounds, tested them for neurogenic properties in ES-derived neural precursor cells, assayed their ability to protect from the amyloid toxicity associated with AD, and determined their metabolic stability. All of the year one milestones we met and we now have the required minimum of six compounds to move into year two studies. In addition, we have made a good start on the work for year two in that some pharmacokinetics and safety studies has been completed.
  • This work will optimize the chances for its true therapeutic potential in AD, and presents a unique opportunity to expand the use of hESCs for the development of a therapeutic for a disease for which there is no cure. This work could lead to a paradigm shift in the treatment of neurodegenerative disease.
  • Introduction: Over 6 million people in the US suffer from Alzheimer’s disease (AD). There are no drugs that prevent the death of nerve cells in AD, nor has any drug been identified that can stimulate their replacement. Even if nerve cells could be replaced, the toxic environment of the brain will kill them unless they are protected by a drug. Therefore, drugs that stimulate the generation of new neurons (neurogenesis) alone will not be effective; a drug with both neurogenic and neuroprotective properties is required. With the ability to use cells derived from human embryonic stem cells (hESCs) as a screen to identify neurogenic compounds, we have shown that it is now be possible to identify and tailor drugs for therapeutic use in AD. This was the overall goal of this application, and to date we have made outstanding progress, making a drug that is both neurogenic for human cells and has therapeutic efficacy in a rigorous mouse model of AD.
  • Year 2 Progress: Using a novel drug discovery paradigm based upon human stem cell derived nerve precursor cells, we have made a very potent drug called CAD-31. CAD-31 potently stimulates neurogenesis in human cells in culture and in mice, and prevents nerve cell death in cell culture models of toxicities associated with old age and AD. Very few, if any, drugs or drug candidates are both neuroprotective and neurogenic, particularly in animals. In the first year of this project, we harnessed the power of hESCs and medicinal chemistry to develop CAD-31. All of the Year 1 milestones were met. In Year 2 we completed all of the required pharmacokinetics and safety studies on the six best compounds synthesized in Year 1. Of those six, one compound, CAD-31, was the best in terms of medicinal chemical, pharmacokinetic, neuroprotective and neurogenic properties. This compound underwent extensive testing for safety and passed with flying colors. It was then put into an AD mouse model where it stimulated neurogenesis, prevented behavioral deficits and some of the disease pathology. All Year 2 milestones were completed. In Year 3 of the project we will determine if CAD-31 is able to reverse AD symptoms in old AD mice that already have the disease. This is the most clinically relevant model of AD since therapies can only be initiated once the disease is identified.
  • This work has produced a novel AD drug candidate that is developed based upon a set of assays never before used by pharmaceutical companies. It presents a unique opportunity to expand the use of hESCs for the development of a therapeutic for a disease for which there is no cure. This work could lead to a paradigm shift in drug discovery for the treatment of neurodegenerative disease.

Use of human iPSC-derived neurons from Huntington’s Disease patients to develop novel, disease-modifying small molecule structural corrector drug candidates targeting the unique, neurotoxic conformation of mutant huntingtin

Funding Type: 
Early Translational IV
Grant Number: 
TR4-06847
Investigator: 
ICOC Funds Committed: 
$1 333 795
Disease Focus: 
Huntington's Disease
Neurological Disorders
Stem Cell Use: 
iPS Cell
oldStatus: 
Active
Public Abstract: 
The long-term objective of this project is to develop a drug to treat Huntington’s disease (HD), the most common inherited neurodegenerative disorder. Characterized by involuntary movements, personality changes and dementia, HD is a devastatingly progressive disease that results in death 10–20 years after disease onset and diagnosis. No therapy presently exists for HD; therefore, this project is highly innovative and ultimately aims to deliver something transformative for the HD patient population. The specific goal of the proposed research will be to achieve preclinical proof-of-concept with a novel small molecule that binds to and ameliorates the neurotoxicity of the mutant huntingtin (mHtt) protein that causes HD. Rationale for development of such compounds comes from previous research that found that mHtt assumes a shape that is selectively toxic to neurons, and that small molecules that disrupt this shape can reduce mHtt’s toxicity in primary neurons. Critical to the proposed studies will be assays that employ human striatal neurons derived from adult and juvenile HD patients and generated with induced pluripotent stem cell (iPSC) technology. These HD i-neurons display many characteristics that are also observed in striatal neurons of HD patients, including reduced survival times. They provide the most genetically precise preclinical system available to test for both drug efficacy and safety.
Statement of Benefit to California: 
The long-term objective of this project is to develop a first-in-class, disease-modifying drug to treat Huntington’s disease (HD), a devastatingly progressive genetic disorder that results in death 10–20 years after disease onset and diagnosis. No therapy presently exists for HD; therefore, this highly innovative project aims to deliver a medical breakthrough that will provide significant benefit for California’s estimated > 2000 HD patients and the family members, friends and medical system that care for them. The proposed research will be performed at a biotechnology startup, a leading academic research center and two contract research organizations, all of which are California-based. The work will over time involve more than 10 California scientists, thereby helping to employ tax-paying citizens and maintain the State’s advanced technical base. Finally, an effective, proprietary drug for the treatment of HD is expected to be highly valuable and to attract favorable financial terms upon out-licensing for development and commercialization. These revenues would flow to the California companies and institutions (including CIRM) that would have a stake in the proceeds.
Progress Report: 
  • The long-term objective for this project was to develop a first-in-class, disease-modifying drug to treat Huntington's disease (HD). This drug would comprise a small molecule that binds to and ameliorates the neurotoxicity of the mutant huntingtin protein (mHtt) that causes HD.
  • The goal of the research conducted under the CIRM Award was to demonstrate development candidate feasibility in vitro with a novel small molecule mHtt detoxifier early lead compound that is potent and efficacious in neurons from HD patients generated using stem cell technology (HD i-neurons) as well as suitable for use in mice as experimental models for HD.
  • The original project strategy was to 1) acquire or synthesize new samples of compounds identified as potential mHtt detoxifiers in the screening campaign conducted 7 years ago; 2) establish or re-establish the cell-free and cultured neuron biological assays needed to characterize potential small molecule mHtt detoxifiers (this work was carried out in the laboratory of our collaborator, Dr. Steven Finkbeiner of the J. David Gladstone Institutes); 3) acquire or synthesize new/novel analogs of the initial hits; 4) test new/novel compounds for activity in a cell-free assay for potential mHtt detoxifier activity; 5) test hits for efficacy in HD and non-HD i-neurons; and 6) profile the in vitro and in vivo pharmacokinetics and absorption, distribution, metabolism and elimination (PK/ADME) profiles of compounds that displayed selective neuroprotection toward HD i-neurons.
  • Specific achievements of the first year of the Project include:
  • • Acquiring 205 previously identified hits or analogs thereof from commercial sources;
  • • Synthesizing an additional 84 novel, designed analogs;
  • • Generating the reagents, re-establishing and implementing the screening assay;
  • • Testing all compounds acquired or synthesized in the screening assay;
  • • Establishing a counterscreen for false positives in the screening assay;
  • • Preliminary screening 48 previously reported hits in the counterscreen;
  • • Testing 14 previously or newly identified hits side-by-side in full concentration-response assays in both the screening and counterscreening assays;
  • • Profiling 11 diverse hits in in vitro PK/ADME assays;
  • • Testing 17 compounds for their ability to ameliorate neurotoxicity in a rodent primary neuron model; and
  • • Preliminary testing 2 previously identified hits in human HD i-neurons.
  • Unfortunately and surprisingly, we observed that all compounds displayed essentially identical profiles in full concentration-response studies in both the screening and counterscreening assays. We interpret this result to indicate that these compounds and structurally related compounds that we considered to be most promising and tested do not in fact bind to mHtt, i.e., they are all false positives. Since no valid starting points exist for continued work, the Project will be terminated after the first award period.

Development of Novel Autophagy Inducers to Block the Progression of and Treat Amyotrophic Lateral Sclerosis (ALS) and Other Neurodegenerative Diseases

Funding Type: 
Early Translational IV
Grant Number: 
TR4-06693
ICOC Funds Committed: 
$2 278 080
Disease Focus: 
Amyotrophic Lateral Sclerosis
Neurological Disorders
Stem Cell Use: 
iPS Cell
oldStatus: 
Active
Public Abstract: 
ALS is a progressive neurodegenerative disease that primarily affects motor neurons (MNs). It results in paralysis and loss of control of vital functions, such as breathing, leading to premature death. Life expectancy of ALS patients averages 2–5 years from diagnosis. About 5,600 people in the U.S. are diagnosed with ALS each year, and about 30,000 Americans have the disease. There is a clear unmet need for novel ALS therapeutics because no drug blocks the progression of ALS. This may be due to the fact that multiple proteins work together to cause the disease and therapies targeting individual toxic proteins will not prevent neurodegeneration due to other factors involved in the ALS disease process. We propose to develop a novel ALS therapy involving small molecule drugs that stimulate a natural defense system in MNs, autophagy, which will remove all of the disease-causing proteins in MNs to reduce neurodegeneration. We previously reported on a class of neuronal autophagy inducers (NAIs) and in this grant will prioritize those drugs for blocking neurodegeneration of human iPSC derived MNs from patients with familial and sporadic ALS to identify leads that will then be tested for efficacy in vivo in animal models of ALS to select a clinical candidate. Since all of our NAIs are FDA approved for treating indications other than ALS, our clinical candidate could be rapidly transitioned to testing for efficacy and safety in treating ALS patients near the end of this grant.
Statement of Benefit to California: 
Neurodegenerative diseases such as ALS as well as Alzheimer’s (AD), Parkinson’s (PD) and Huntington’s Disease (HD) are devastating to the patient and family and create a major financial burden to California (CA). These diseases are due to the buildup of toxic misfolded proteins in key neuronal populations that leads to neurodegeneration. This suggests that common mechanisms may be operating in these diseases. The drugs we are developing to treat ALS target this common mechanism, which we believe is an impairment of autophagy that prevents clearance of disease-causing proteins. Effective autophagy inducers we identify to treat ALS may turn out to be effective in treating other neurodegenerative diseases. This could have a major impact on the health care in CA. Most important in our studies is the translational impact of the use of patient iPSC-derived neurons and astrocytes to identify a new class of therapeutics to block neurodegeneration that can be quickly transitioned to testing in clinical trials for treating ALS and other CNS diseases. Future benefits to CA citizens include: 1) development of new treatments for ALS with application to other diseases such as AD, HD and PD that affect thousands of individuals in CA; 2) transfer of new technologies to the public realm with resulting IP revenues coming into the state with possible creation of new biotechnology spin-off companies and resulting job creation; and 3) reductions in extensive care-giving and medical costs.
Progress Report: 
  • ALS is a progressive neurodegenerative disease that primarily affects motor neurons (MNs). It results in paralysis and loss of control of vital functions, such as breathing, leading to premature death. Much scientific evidence indicates that ALS is due to the buildup of toxic misfolded proteins in key neuronal populations that leads to neurodegeneration. In this CIRM-funded project, we are developing drugs that can improve a cellular process called “autophagy” by which cells, including neurons, clear out built-up toxic misfolded proteins and increase their longevity. We had discovered that a series of FDA-approved drugs already on the market for other indications happen to induce autophagy in a manner that is independent of their original purpose. Our goal is to show that these FDA-approved drugs can induce autophagy and slow neurodegeneration in ALS patient-derived neurons, and to repurpose these drugs for ALS. In the last year, we have made significant progress towards testing these drugs on neurons that we derived from induced pluripotent stem cells engineered from skin cells taken from ALS patients. We have built robotic microscopes that can rapidly image ALS patient neurons that are treated with drugs in the lab and determine whether any of these “autophagy-inducing” FDA-approved drugs slowdown the rate of neurodegeneration. We have optimized large-scale methods to grow patient neurons, treat them with drugs, image them over many days, and analyze the images to measure neurodegeneration. In August 2014, we published a paper in the journal Nature Chemical Biology that showed two FDA-approved drugs can in fact induce autophagy and increase the clearing of an ALS-related protein called TDP43 in neurons. The drugs were also able to slow neurodegeneration in neurons and astrocytes derived from a familial ALS patient with an altered version of the TDP43 gene. We have now obtained stem cells from broader types of familial ALS as well as sporadic ALS patients, have made neurons from their stem cells, and have treated their stem cell-derived neurons with more than 10 autophagy-inducing drugs at varying concentrations to determine whether autophagy-inducers can slow neurodegeneration in neurons from broader forms of ALS. These neurons are currently being imaged using our robotic microscope. In addition, we have started to make astrocytes from patient stem cells and plan to test the drugs on astrocytes in the coming months.

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