Neurological Disorders

Coding Dimension ID: 
303
Coding Dimension path name: 
Neurological Disorders
Funding Type: 
Basic Biology V
Grant Number: 
RB5-07011
Investigator: 
ICOC Funds Committed: 
$1 161 000
Disease Focus: 
Alzheimer's Disease
Neurological Disorders
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Closed
Public Abstract: 

We propose to elucidate pathways of genes that lead from early causes to later defects in Alzheimer’s Disease (AD), which is common, fatal, and for which no effective disease-modifying drugs are available. Because no effective AD treatment is available or imminent, we propose to discover novel genetic pathways by screening purified human brain cells made from human reprogrammed stem cells (human IPS cells or hIPSC) from patients that have rare and aggressive hereditary forms of AD. We have already discovered that such human brain cells exhibit an unique biochemical behavior that indicates early development of AD in a dish. Thus, we hope to find new drug targets by using the new tools of human stem cells that were previously unavailable. We think that human brain cells in a dish will succeed where animal models and other types of cells have thus far failed.

Statement of Benefit to California: 

Alzheimer’s Disease (AD) is a fatal neurodegenerative disease that afflicts millions of Californians. The emotional and financial impact on families and on the state healthcare budget is enormous. This project seeks to find new drug targets to treat this terrible disease. If we are successful our work in the long-term may help diminish the social and familial cost of AD, and lead to establishment of new businesses in California using our approaches.

Progress Report: 
  • The goal of this project has been to understand how neurons made from stem cells that are genetically engineered to develop Alzheimer's disease in a dish generate abnormal biochemistry that we can measure with simple assays. In the first year of this project we developed new probes for the pathway we are trying to measure. However, we encountered technical obstacles that interfere with our ability to evaluate the function of this pathway. We think we have identified the cause of the problems and in the second year of the project we will initiate experiments to solve these problems and rigorously evaluate how genetic mutations that cause abnormal Alzheimer's biochemistry generate the abnormal biochemistry in our human neural system made from stem cells.
Funding Type: 
Early Translational from Disease Team Conversion
Grant Number: 
TRX-01471
Investigator: 
ICOC Funds Committed: 
$4 139 754
Disease Focus: 
Amyotrophic Lateral Sclerosis
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 
Statement of Benefit to California: 
Progress Report: 
  • Project Description and Rationale:
  • Amyotrophic Lateral Sclerosis (ALS) is the most common adult motor neuron disease, affecting 30,000 people in the US and the typical age of onset is in the mid-50s or slightly younger. ALS is a degenerative neural disease in which the damage and death of neurons results in progressive loss of the body’s functions until death, which is usually in 3-5 years of diagnosis. Current ALS treatments are primarily supportive, and providing excellent clinical care is essential for patients with ALS; however, there is an urgent need for treatments that significantly change the disease course. The only Food and Drug Administration approved, disease-specific medication for treatment of ALS is Rilutek (riluzole); which demonstrated only a modest effect on survival (up to 3 months) in clinical trials.
  • The ALS Disease Team/Early Translational project is focused on developing an ALS therapy based on human embryonic stem cell (ESC) derived neural stem cells (NSC) and/or astrocyte precursor cells transplanted into the ventral horn of the spinal cord. Several lines of evidence strongly support the approach of transplanting cells that exhibit the capacity to migrate, proliferate and mature into normal healthy astrocytes which can provide a neuroprotective effect for motor neurons and reduce or prevent neural damage and disease progression in ALS. Strong evidence has been generated from extensive studies in culture dishes and in animal models to support the concept that providing normal astrocytes in the proximity of α-motor neurons can protect them from neural damage.
  • Project Plan and Progress:
  • Multiple ESC lines were acquired in 2 rounds based on early and later availability. The first round of ESCs included ESCs from City of Hope (GMP H9) and the University of California, San Francisco (UCSF4). The second round included ESCs from the University of California, San Francisco [UCSFB6 (aka UCSF4.2) and UCSFB7 (aka UCSF4.3)] and from BioTime (ESI-017). These ESC lines were tested for their ability to survive and expand under conditions required for producing a cellular therapy (FDA GMP-like and GTP compliant conditions). From these ESC lines, NSCs were generated, expanded and characterized to determine their ability to produce stable and consistent populations of NSCs under conditions required for producing a cellular therapy.
  • For the first round of cell lines, both UCSF4 and H9 were successfully induced to produce NSCs, which were mechanically enriched, expanded and implanted into immunodeficient rats and a rat model of ALS (SOD1G93A). For this small-scale in vivo screen, implanted UCSF4 and H9 NSCs survived, migrated and differentiated into neurons and astrocytic cells in 3-5 weeks, without producing tumors or other unwanted structures. NSCs from both UCSF4 and H9 performed similarly in culture and in vivo, thus the decision to use UCSF4 in the larger-scale in vivo studies for safety (implant into immunodeficient rats) and efficacy/proof of concept (SOD1G93A ALS model rats) was weighted by the difficulties obtaining H9 for future studies for a therapeutic product. These larger-scale studies began August 2013 (earlier than projected), with expected completion in February 2014.
  • For the second round of ESC lines (UCSFB6, UCSFB7 and BioTime ESI-017), UCSFB6 and UCSFB7 ESCs expanded well, while ESI-017 expansion was less robust. Because UCSFB6 and UCSFB7 ESCs are from the same blastomere, we decided to continue to NSC production with only UCSFB7, keeping UCSFB6 in reserve as a back-up. UCSFB7 ESCs were successfully induced to produce NSCs, which were mechanically enriched, expanded and implanted into immunodeficient rats and a rat model of ALS (SOD1G93A). The results from these studies are pending (some animals are still in-life), but early histology suggests the cell survival is similar to UCSF4 and H9. A second round of large-scale in vivo studies is planned to start January 2014 to evaluate this NSC line. By September 2014, the “best” NSC line will be selected as a therapeutic candidate for definitive pre-clinical studies and entry into clinical trials.
  • ESC production under GMP-like condition has been completed at the UC Davis GMP facility. UC Davis generated the first batch of NSCs, which were not sufficiently homogeneous for successful expansion beyond approximately passage 10. This prompted UCSD to investigate multiple enrichment strategies, which were tested on multiple cell lines to ensure method reproducibility. A mechanical enrichment method reproducibly resulted in more homogeneous NSC cultures, capable of expansion for 20 – 30 passages, or more. The NSC generation and enrichment methods are currently being transferred to UC Davis and the UCSD scientist who developed the methods will work side-by-side with the UC Davis GMP production team to ensure successful method transfer to the GMP facility.
  • UCSF4 NSCs are also in use in a CIRM supported early translation study for spinal cord injury.
  • Project Description and Rationale:
  • Amyotrophic Lateral Sclerosis (ALS) is the most common adult motor neuron disease, affecting 30,000 people in the US and the typical age of onset is in the mid-50s or slightly younger. ALS is a degenerative neural disease in which the damage and death of neurons results in progressive loss of the body’s functions until death, which is usually in 3-5 years of diagnosis. Current ALS treatments are primarily supportive, and providing excellent clinical care is essential for patients with ALS; however, there is an urgent need for treatments that significantly change the disease course. The only Food and Drug Administration approved, disease-specific medication for treatment of ALS is Rilutek (riluzole); which demonstrated only a modest effect on survival (up to 3 months) in clinical trials.
  • The ALS Disease Team/Early Translational project is focused on developing an ALS therapy based on human embryonic stem cell (ESC) derived neural stem cells (NSC) and/or astrocyte precursor cells transplanted into the ventral horn of the spinal cord. Several lines of evidence strongly support the approach of transplanting cells that exhibit the capacity to migrate, proliferate and mature into normal healthy astrocytes which can provide a neuroprotective effect for motor neurons and reduce or prevent neural damage and disease progression in ALS.
  • Year 2 Progress Summary:
  • The longer-term, larger-scale in vivo safety and efficacy studies using the lines that showed the most promise during previous screening studies (UCSF4 and ESI-017 NSCs) have been completed. The safety studies were performed in immunodeficient rats to evaluate the survival, migration, differentiation, function and tumorigenicity of implanted NSCs at 3 weeks, 2 months and 6 months post implant. The efficacy studies were conducted in a transgenic SOD1G93A ALS rat model to evaluate safety and cell fate in the background of disease, as well as, to evaluate disease-modifying activity (e.g. neural protection/proof-of-concept) of the implanted NSCs.
  • NOTE: A labeling error occurred during expansion and banking of the ESCs at UC Davis, and the cell line labeled as UCSFB7 (aka UCSF4.3) was determined by DNA fingerprinting to actually be ESI-017. Previous NSC generation, characterization and in vivo screening data was reported for cell line UCSFB7 (aka UCSF4.3), which was actually for ESI-017.
  • Both UCSF4 and ESI-017 NSCs were deemed acceptable in 2 out of 3 of the minimal acceptance criteria:
  • 1) Long-term survival in nude and SOD1G93A rats
  • 2) No formation of tumors or other unwanted structures when implanted into nude or SOD rats.
  • The third criterion: at least 10% greater α-motor neuron counts in cell-injected animals as compared to medium injected controls (or cell-injected side compared to the non-injected, or contralateral side) was not met due to a) variability of α-motor neuron counts and b) the aggressive nature of the current SOD1G93A rat ALS model and resulting very short 2 month treatment window which exceeds the length of time for the migration, expansion, differentiation and maturation of sufficient astrocytes to provide a neural protective effect in all implanted animals.
  • UCSF4 NSCs were originally selected as the developmental candidate, however, there are compelling reasons to reconsider ESI-017 NSCs: 1) UC Davis has found ESI-017 NSCs relatively easy to generate and is having difficulty generating UCSF4 NSCs; and 2) recent hisotological evaluations suggest that ESI-017 NSCs produce mature astrocytes earlier in vivo than UCSF4 NSCs. We are working with UC Davis on generation of UCSF4 NSCs and are quantifying astrocyte maturation histology (e.g. GFAP) to make a well-supported developmental candidate selection.
  • In parallel, mRNA sequencing has been performed 1) on cells produced in the course of this project to identify potential markers predictive of in vivo fate, 2) on naïve SOD1G93A rats to explore markers of disease onset and progression that could potentially be used as surrogate markers of disease modulation in place of motor neuron counts, and 3) on NSCs implanted into nude and SOD1G93A rats to identify potential markers of long-term post-transplant NSC cell fate and host response.
  • Project Description and Rationale:
  • Amyotrophic Lateral Sclerosis (ALS) is the most common adult motor neuron disease, affecting 30,000 people in the US and the typical age of onset is in the mid-50s or slightly younger. ALS is a degenerative neural disease in which the damage and death of neurons results in progressive loss of the body’s functions until death, which is usually in 3-5 years of diagnosis. Current ALS treatments are primarily supportive, and providing excellent clinical care is essential for patients with ALS; however, there is an urgent need for treatments that significantly change the disease course. The only FDA approved, disease-specific medication for treatment of ALS is Rilutek (riluzole); which demonstrated only a modest effect on survival (up to 3 mo.) in clinical trials.
  • The ALS Disease Team/Early Translational project is focused on developing an ALS therapy based on human embryonic stem cell (ESC) derived neural stem cells (NSC) and/or astrocyte precursor cells transplanted into the ventral horn of the spinal cord. Several lines of evidence strongly support the approach of transplanting cells that exhibit the capacity to migrate, proliferate and mature into normal astrocytes and provide a neuroprotection for motor neurons to reduce/prevent neural damage and disease progression.
  • Year 3 (6 month) Progress Summary:
  • NSCs generated with clonal enrichment from ESI-017 ESCs have a similar capacity to stably expand in vitro; and to survive, migrate and differentiate into neuronal and astrocytic cells in vivo without generating teratomas or other unwanted tissue formations when implanted into nude or SOD1 ALS rats. UCSF4 NSCs were originally selected as the developmental candidate, however, we changed to ESI-017 NSCs because: 1) UC Davis found ESI-017 NSC generation relatively easy but were unable to produce UCSF4 NSCs even with several method modifications; and 2) histology suggests that ESI-017 NSCs produce mature astrocytes earlier than UCSF4 NSCs.
  • ESI-017 NSCs generated at UC Davis under research conditions with predominantly GMP compatible reagents were implanted into athymic rats in order to compare the in vivo fate to NSCs generated at UCSD. Animals were perfused 2 months and 6 months post implant. Histology showed graft survival and differentiation of implanted ESI-017 NSCs generated at UCSD and UC Davis are similar, and further confirm successful transfer of NSC production methods the UC Davis.
  • The aggressive disease presentation of the current SOD1 rat ALS model results in a very short 2-month treatment window which exceeds the length of time for the migration, expansion, differentiation and maturation of sufficient astrocytes to provide a neural protective effect; and the variability of α-motor neuron counts precludes the use of this animal model demonstration of efficacy or proof-of-concept. We discussed these issues during a "pre-pre-IND" call with CBER/OCTGT/FDA on September 9, 2014. Prior to the call, Mercedes Serabian, Chief, Pharmacology/Toxicology Branch provided informal general comments which included examples of POC study endpoints were provided (e.g. motor neuron counts, levels of glutamate transport, electrophysiology/neurophysiology, etc.). During the call, discussion regarding glutamate excitotoxicity in ALS, and demonstrating that our heNSCs (or glial progeny) have the capacity to preserve/replace lost glutamate transporter activity in a model of ALS was “on the table” as potentially acceptable demonstration of POC, and we should have another pre-pre-IND call when we have such data.
  • In order to generate POC data for further discussion with the FDA we preformed additional histology on existing spinal cord tissue and initiated a collaboration with Don Cleveland’s and Brian Kaspar’s labs to perform in vitro co-culture experiments using ESI-017 NSCs generated at UCSD.
  • Histology: additional histological evaluation of spinal cord tissue from ESI-017 implanted nude and SOD1 rats demonstrated GLAST expression in grafted human astrocytes, suggesting active glutamate buffering activity.
  • In vitro co-culture experiments: ESI-017 NSCs generated at UCSD were shipped to Brian Kaspar’s lab where they were expanded and differentiated into astrocytes using their published methods. Human astrocytes were co-cultured with GFP positive motor neurons (MN) and at various time during culture, images were recorded and processed for survival cell counts and neurite length measurements. After 5 days of co-culture, astrocytes generated from ESI-017 NSCs provided motor neuron support similar to that provided by the normal control astrocytes whereas astrocytes derived from NSCs isolated post mortem from spinal cord tissue of patients with either familial ALS (FALS) or sporadic ALS (SALS) were toxic to motor neurons in co-culture. This co-culture experiment was repeated with similar findings (data analysis is in progress).
  • Collaboration with the Kaspar lab will continue beyond the end of this award.
Funding Type: 
Tissue Collection for Disease Modeling
Grant Number: 
IT1-06571
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$530 265
Disease Focus: 
Autism
Neurological Disorders
Pediatrics
oldStatus: 
Active
Public Abstract: 

Autism spectrum disorders (ASD) are a family of disabling disorders of the developing brain that affect about 1% of the population. Studying the biology of these conditions has been difficult as they have been challenging to represent in animal models. The core symptoms of ASD, including deficits in social communication, imagination and curiosity are intrinsically human and difficult to model in organisms commonly studied in the laboratory. Ideally, the mechanisms underlying ASDs need to be studied in human patients and in their cells. Since they maintain the genetic profile of an individual, studying neurons derived from human induced pluripotent stem cells (hiPSC) is attractive as a method for studying neurons from ASD patients. hiPSC based studies of ASDs hold promise to uncover deficits in cellular development and function, to evaluate susceptibility to environmental insults, and for screening of novel therapeutics. In this project our goal is to contribute blood and skin samples for hiPSC research from 200 children with an ASD and 100 control subjects to the CIRM repository. To maximize the value of the collected tissue, all subjects will have undergone comprehensive clinical evaluation of their ASD. The cells collected through this project will be made available to the wider research community and should result in a resource that will enable research on hiPSC-derived neurons on a scale and depth that is unmatched anywhere else in the world.

Statement of Benefit to California: 

The prevalence and impact of Autism Spectrum Disorders (ASD) in California is staggering. California has experienced 13% new ASD cases each year since 2002. ASD are a highly heritable family of complex neurodevelopmental conditions affecting the brain, with core symptoms of impaired social skills, language, behavior and intellectual abilities. The majority with an ASD experience lifelong disability that requires intensive parental, school, and social support. The result has been a 12-fold increase in the number of people receiving ASD services in California since 1987, with over 50,000 people with ASDs served by developmental and regional centers. Within the school system, the number of special education students with ASD in California has more than tripled between 2002 and 2010. The economic, social and psychological toll is enormous.
It is critical to both prevent and develop effective treatments for ASD. While rare genetic mutations account for a minority of cases, our understanding of idiopathic ASD (>85% of cases) is extremely limited. Mechanisms underlying ASDs need to be studied in human patients and in cells that share the genetic background of these patients. Since they maintain the complete genetic background of an individual, hiPSCs represent a very practical and direct method for investigating neurons from ASD patients to uncover cellular deficits in their development and function, and for screening of novel therapeutics.

Progress Report: 
  • Autism Spectrum Disorders (ASD) have a worldwide prevalence of 1% (>1.5 million in the US) and a lifetime cost per affected individual of $3.2M. ASDs are amongst the most heritable of psychiatric disorders. Genome Wide Association studies utilizing samples in the thousands provide only weak evidence for common allele risk effects; positive findings rarely replicate, and genetic effects sizes are small (odds ratios of ~1.1). In contrast, evidence to date for risk or causation conferred by rare variation, particularly rare copy number variants, is very strong. Pathway analyses of the rare mutations implicated and genome-wide transcriptome analysis of brain and blood tissue provide converging evidence that neural-related pathways are central to the development of autism. Core impairments of ASDs, such as imagination and curiosity about the environment, cannot be modeled well in other organisms. The mechanisms underlying ASDs need to be studied in humans and cells that share the genetic background of the patients, such as neurons from patients derived from induced pluripotent cell lines (iPSC).
  • Our goal was to provide the CIRM repository with samples from 200 well characterized individuals with an ASD and 100 demographically matched controls. To date we have enrolled 63 participants.
Funding Type: 
Basic Biology IV
Grant Number: 
RB4-05855
Investigator: 
ICOC Funds Committed: 
$1 387 800
Disease Focus: 
Neuropathy
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 

The use of stem cells or stem cell-derived cells to treat disease is one important goal of stem cell research. A second, important use for stem cells is the creation of cellular models of human development and disease, critical for uncovering the molecular roots of illness and testing new drugs. However, a major limitation in achieving these goals is the difficulty in manipulating human stem cells. Existing means of generating genetically modified stem cells are not ideal, as they do not preserve the normal gene regulation, are inefficient, and do not permit removal of foreign genes.

We have developed a method of genetically modifying mouse embryonic stem cells that is more efficient than traditional methods. We are adapting this approach for use with human embryonic stem cells, so that these cells can be better understood and harnessed for modeling, or even treating, human diseases. We will use this approach to create a human stem cell model of Charcot-Marie-Tooth (CMT) disease, an inherited neuropathy. How gene dysfunction leads to nerve defects in CMT is not clear, and there is no cure or specific therapy for this neurological disease. Thus, we will use our genetic tools to investigate how gene function is disrupted to cause CMT. By developing these tools and using them to gain understanding of CMT, we will illustrate how this system can be used to gain insight into other important diseases.

Statement of Benefit to California: 

Although human stem cells hold the potential to generate new understanding about human biology and new approaches to important diseases, the inability to efficiently and specifically modify stem cells currently limits the pace of research. Also, there is presently no safe means of changing genes compatible with the use of the stem cells in therapies. We are developing new genetic tools to allow for the tractable manipulation of human stem cells. By accelerating diverse other stem cell research projects, these tools will enhance the scientific and economic development of California.

We will use these tools to create cellular models of Charcot-Marie-Tooth (CMT), a neurological disease with no cure that affects about 15,000 Californians. This model will facilitate understanding of the etiology of CMT, and may lead to insights that can be used to develop specific therapies.

Beyond gaining insight into CMT, the ability to engineer specific genetic changes in human stem cells will be useful for many applications, including the creation of replacement cells for personalized therapies, reporter lines for stem cell-based drug screens, and models of other diseases. Thus, our research will assist the endeavors of the stem cell community in both the public and private arenas, contributing to economic growth and new product development. This project will also train students and postdoctoral scholars in human stem cell biology, who will contribute to the economic capacity of California.

Progress Report: 
  • An important use for stem cells is the creation of cellular models of human development and disease, critical for uncovering the molecular roots of illness and testing new drugs. However, a major limitation in achieving these goals is the difficulty in manipulating human stem cells. We have developed a method of genetically modifying mouse embryonic stem cells that is more efficient than traditional methods. During the first year of this project, we adapted this approach for use with human embryonic stem cells. We have also created gene trap mutations in a diversity of human embryonic stem cell genes that can be used to better harness human embryonic stem cells for modeling, or even treating, human diseases.
  • An important use for stem cells is the creation of cellular models of human development and disease, critical for uncovering the molecular roots of illness and testing new drugs. However, a major limitation in achieving these goals is the difficulty in manipulating human stem cells. We have developed a method of genetically modifying mouse embryonic stem cells that is more efficient than traditional methods. During the second year of this project, we took advantage of new methods using the CRISPR/Cas9 system to develop novel approaches to modifying human embryonic stem cells. We have also created reversible gene trap mutations in a diversity of human embryonic stem cell genes that can be used to better harness human embryonic stem cells for modeling, or even treating, human diseases.
Funding Type: 
Basic Biology IV
Grant Number: 
RB4-06045
Investigator: 
Name: 
Type: 
PI
ICOC Funds Committed: 
$1 393 200
Disease Focus: 
Amyotrophic Lateral Sclerosis
Neurological Disorders
Dementia
Neurological Disorders
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 

Amyotrophic lateral sclerosis (ALS) is an idiopathic adult-onset degenerative disease characterized by progressive weakness from loss of upper and lower motor neurons. Onset is insidious, progression is essentially linear, and death occurs within 3-5 years in 90% of patients. In the US, 5,000 deaths occur per year and in the world, 100,000. In October, 2011, the causative gene defect in a long sought after locus on chromosome 9 for ALS, frontotemporal dementia (FTD) and overlap ALS-FTD was identified to be a expansion of a hexanucleotide repeat in the uncharacterized C9ORF72 gene. The goal of the proposed research is to generate human stem cell models from cells derived from ALS patients with the C9ORF72 expanded repeats and relevant control cells using genome-editing technology. We will also generate a stem cell model expressing the repeat independent of the C9ORF72 gene to study if the repeat alone is causing neural defects. Using advanced genome technologies, biochemical and cellular approaches, we will study the molecular pathways affected in motor neurons derived from these stem cell models. Finally, we will use innovative technologies to rescue the abnormal phenotypes that arise from the expanded repeat in human motor neurons. Completion of the proposed research is expected to transform our understanding of the regulatory and pathogenetic mechanisms underlying ALS and FTD, and establish therapeutic options for these debilitating diseases.

Statement of Benefit to California: 

Our research provides the foundation for decoding the mechanisms that underlie the single most frequent genetic mutation found to contribute to both ALS and FTD, debilitating neurological diseases that impact many Californians. In California, the expected prevalence of ALS (the number of total existing cases) is 2,200 to 3,000 cases at any one time, and the incidence is 750-1,100 new cases each year. The number of FTD cases is five times as many. Our research has and will continue to serve as a basis for understanding deviations from normal and disease patient neuronal cells, enabling us to make inroards to understanding neurological disease modeling using neurons differentiated from reprogammed patient-specific lines. Such disease modeling will have great potential for California health care patients, pharmaceutical and biotechnology industries in terms of improved human models for drug discovery and toxicology testing. Our improved knowledge base will support our efforts as well as other Californian researchers to study stem cell models of neurological disease and design new diagnostics and treatments, thereby maintaining California's position as a leader in clinical research.

Progress Report: 
  • Expanded hexanucleotide repeats in the C9ORF72 gene were identified in Oct 2011 as a cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), thus identifying the single most frequent genetic cause of each and connecting them to repeat expansion disease. We are developing stem cell disease models to enable key hypotheses of pathogenesis and new interventions to be tested. We have successfully engineered stem cell models to analyze the effects of C9ORF72 mutations, and have differentiated these stem cell models into motor neurons which enabled us to conduct transcriptomic and biochemical studies. In addition, we have utilized antisense-oligonucleotides (ASOs) from ISIS Pharmaceuticals to deplete mutant C9ORF72 in motor neurons. We expect our efforts to provide mechanistic insights and a potential therapy in human cells.
  • In this period, we have generated C9ORF72 induced pluripotent stem cells and differentiated them into mature motor neurons. We have found that expression profiles from previous reports are largely irreproducible, suggesting there is substantial heterogeneity in the cells from patients. To address the issue of cell-type specificity, we have developed cell-type specific reporters in these lines and have generated astrocytes and motor neurons.
Funding Type: 
Basic Biology IV
Grant Number: 
RB4-05886
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$1 392 426
Disease Focus: 
Neurological Disorders
Stem Cell Use: 
Directly Reprogrammed Cell
oldStatus: 
Active
Public Abstract: 

Many human diseases and injuries that affect the brain and nervous system could potentially be treated by either introducing healthy neurons or persuading the cells that normally provide supporting functions to become functioning neurons. A number of barriers must be traversed to bring these goals to practical therapies. Recently our laboratory and others have found ways of converting different human cell types to functioning neurons. Surprisingly, two routes for the production of neurons have been discovered. Our preliminary evidence indicates that these two routes are likely to work together and therefore more effective ways of producing neurons can likely be provided by understanding these two routes, which is one aim of this application. Another barrier to effective treatment of human neurologic diseases has been the inability to develop good models of human neurologic disease due to inability to sample tissues from patients with these diseases. Hence we will understand ways of making neurons from blood cells and other cells, which can be easily obtained from patients with little or no risk. Our third goal will be to understand how different types of neurons can be produced from patient cells. We would also like to understand the barriers and check points that keep one type of cell from becoming another another type of cell. Understanding these mysterious processes could help provide new sources of human cells for replacement therapies and disease models.

Statement of Benefit to California: 

The state of California and its citizens are likely to benefit from the work described in this proposal by the development of more accurate models for the testing of drugs and new means of treatment of human neurologic diseases. Presently these diseases are among the most common afflicting Californians, as well as others and will become more common in an aging population. Common and devastating diseases such as Alzheimer’s, Schizophrenia, Parkinson's Disease, and others lack facile cell culture models that allow one to probe the basis of the disease and to test therapies safely and without risk to the patient. Our work is already providing these models, but we hope to make even better ones by understanding the fundamental processes that allow one cell type (such as a skin cell or blood cell) to be converted to human neurons, where the disease process can be investigated. In the past the inability to make neurons from patients with specific diseases has been a major roadblock to treatment. In the future the studies described here might be able to provide healthy neurons to replace ones loss through disease or injury.

Progress Report: 
  • During the past year, our laboratory has investigated the way that human skin cells can be changed to neurons. To do this, we have used a natural switch that occurs as embryonic cells decide to become neurons. We have found that this process proceeds in a highly ordered series of stages that involve first a resetting of fundamental cell biologic processes characteristic of neurons. This is followed by activation of genes encoding proteins that allow different types of neurons to interact and develop communication between one another. This finding surprised us since we expected to find changes in transcription factors, which instruct the formation of neurons. Instead, we find that the natural switching mechanism in neurons first regulates cell-to-cell communication.
  • We are exploring the way that normal human skin and other types of cells can be converted to neurons. We have found that there are at least two fundamental genetic pathways of doing this that are influenced by different genes and may therefore represent a fertile ground for developing new methods for converting cells of different types to neurons. This could perhaps be useful for replacing neurons from other cell types in states where neurons are damaged or lost such as a variety of neurodegenerative diseases.
Funding Type: 
Basic Biology IV
Grant Number: 
RB4-06345
Investigator: 
ICOC Funds Committed: 
$1 360 450
Disease Focus: 
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 

A subset of intellectual disability cases in humans are caused by mutations in an X-linked gene essential for a quality control mechanism called nonsense-mediated RNA decay (NMD). Patients with mutations in this gene—UPF3B—commonly have not only ID, but also schizophrenia, autism, and attention-deficit/hyperactivity disorder. Thus, the study of UPF3B and NMD may provide insight into a wide spectrum of cognitive and psychological disorders. To examine how mutations in UPF3B can cause mental defects, we will generate and characterize induced-pluripotent stem cells from intellectual disability patients with mutations in the UPF3B gene. In addition to having a role in neural development, our recent evidence suggests that NMD is important for maintaining the identity of ES cells and progenitor cells. How does NMD do this? While NMD is a quality control mechanism, it is also a well characterized biochemical pathway that serves to rapidly degrade specific subsets of normal messenger ribonucleic acids (mRNAs), the transiently produced copies of our genetic material: deoxyribonucleic acid (DNA). We have obtained evidence that NMD preferentially degrades mRNAs that interfere with the stem cell program (i.e., NMD promotes the decay mRNAs encoding proteins that promote differentiation and inhibit cell proliferation). In this proposal, we will identify the target mRNAs of NMD in stem and progenitor cells and directly address the role of NMD in maintaining the stem-like state.

Statement of Benefit to California: 

iPS cells provide a means to elucidate the mechanisms underlying diseases that afflict a growing number of Californians. Our proposed project concerns making and testing iPS cells from patients with mutations in the UPF3B gene, all of whom have intellectual disabilities. In addition, many of these patients have autism, attention-deficit disorders, and schizophrenia. By using iPS cells to identify the cellular and molecular defects in these patients, we have the potential to ultimately ameliorate the symptoms of many of these patients. This is important, as over 1.6 million people in California have serious mental illness. Moreover, a large proportion of patients with UPF3B mutations have autism, a disorder that has undergone an alarming 12-fold increase in California between 1987 and 2007.
The public mental health facilities in California are inadequate to meet the needs of people with mental health disorders. Furthermore, what is provided is expensive: $4.4 billion was spent on public mental health agency services in California in 2006. Mental health problems also exert a heavy burden on California’s criminal justice system. In 2006, over 11,000 children and 40,000 adults with mental health disorders were incarcerated in California’s juvenile justice system.
Our research is also directed towards understanding fundamental mechanisms by which all stem cells are maintained, which has the potential to also impact non-psychiatric disorders suffered by Californians.

Progress Report: 
  • A key quality of stem cells is their ability to switch from a proliferative cell state in which they reproduce themselves to a differentiated cell state that ultimately allows them to carry out the functions of a fully mature cell. Most research on the nature of this switch has focused on the role of proteins that determine whether the genetic material—DNA—generates a copy of it itself in the form of messenger RNA, a process called transcription. In stem cells, such proteins—which are called transcription factors—activate the production of messenger RNAs encoding proteins that promote the proliferative and undifferentiated cell state. They also increase the production of messenger mRNAs that encode inhibitors of differentiation and cell proliferation. The level and profile of such transcription factors are altered in response to signals that trigger stem cells to differentiate. For example, transcription factors that promote the undifferentiated cell state are decreased in level and transcription factors that drive differentiation down a particular lineage are increased in level. While this transcription factor-centric view of stem cells explains some aspects of stem cell biology, it is, in of itself, insufficient to explain many of their behaviors, including both their ability to maintain the stem-like state and to differentiate. We hypothesize that a molecular pathway that complements transcription-base mechanisms in controlling stem cell maintenance vs. differentiation decisions is an RNA decay pathway called nonsense-mediated RNA decay (NMD). Messenger RNA decay is as important as transcription in determining the level of messenger RNA. Signals that trigger increased decay of a given messenger RNA leads to decreased levels of its encoded protein, while signals that trigger the opposite response increase the level of the encoded protein. Our project revolves around two main ideas. First, that NMD promotes the stem-like state by preferentially degrading messenger RNAs that encode differentiation-promoting proteins and proliferation inhibitor proteins. Second, that NMD must be downregulated in magnitude to allow stem cells to differentiate. During the progress period, we obtained substantial evidence for both of these hypotheses. With regard to the first hypothesis, we have used genome-wide approaches to identify hundreds of messenger RNAs that are regulated by NMD in both in vivo (in mice) and in vitro (in cell lines). To determine which of these messenger mRNAs are directly degraded by NMD, we have used a variety of approaches. This work has revealed that NMD preferentially degrades messenger RNAs encoding neural differentiation inhibitors and proliferation inhibitors in neural stem cells. In contrast, very few messenger RNAs encoding pro-stem cell proteins or pro-proliferation proteins are degraded by NMD. Together this provides support for our hypothesis that NMD promotes the stem-like state by shifting the proportion of messenger RNAs in a cell towards promoting an undifferentiated, proliferative cell state. With regard to the second hypothesis, we have found that many proteins that are directly involved in the NMD pathway are downregulated upon differentiation of stem and progenitor cells. Not only are NMD proteins reduced in level, but we find that the magnitude of NMD itself is reduced. We have used a variety of molecular techniques to determine whether this NMD downregulatory response has a role in neural differentiation and found that NMD downreglation is both necessary and sufficient for this event. Such experiments have also revealed particular messenger mRNAs degraded by NMD that are crucial for the NMD downregulatory response to promote neural differentiation. Our research has implications for intellectual disability cases in humans caused by mutations in an X-linked gene essential for NMD. Patients with mutations in this gene—UPF3B—not only have intellectual disability, but also schizophrenia, autism, and attention-deficit/hyperactivity disorder. Thus, the study of NMD may provide insight into a wide spectrum of cognitive and psychological disorders. We are currently in the process of generating induced-pluripotent stem (iPS) cells from intellectual disability patients with mutations in the UPF3B gene towards this goal.
  • A key quality of stem cells is their ability to switch from a proliferative cell state in which they reproduce themselves to a differentiated cell state that ultimately allows them to carry out the functions of a fully mature cell. Most research on the nature of this switch has focused on the role of proteins that determine whether the genetic material—DNA—generates a copy of it itself in the form of messenger RNA, a process called transcription. In stem cells, such proteins—which are called transcription factors—activate the production of messenger RNAs encoding proteins that promote the proliferative and undifferentiated cell state. They also increase the production of messenger mRNAs that encode inhibitors of differentiation and cell proliferation. The level and profile of such transcription factors are altered in response to signals that trigger stem cells to differentiate. For example, transcription factors that promote the undifferentiated cell state are decreased in level and transcription factors that drive differentiation down a particular lineage are increased in level. While this transcription factor-centric view of stem cells explains some aspects of stem cell biology, it is, in of itself, insufficient to explain many of their behaviors, including both their ability to maintain the stem-like state and to differentiate. We hypothesize that a molecular pathway that complements transcription-base mechanisms in controlling stem cell maintenance vs. differentiation decisions is an RNA decay pathway called nonsense-mediated RNA decay (NMD). Messenger RNA decay is as important as transcription in determining the level of messenger RNA. Signals that trigger increased decay of a given messenger RNA leads to decreased levels of its encoded protein, while signals that trigger the opposite response increase the level of the encoded protein. Our project revolves around two main ideas. First, that NMD promotes the stem-like state by preferentially degrading messenger RNAs that encode differentiation-promoting proteins and proliferation inhibitor proteins. Second, that NMD must be downregulated in magnitude to allow stem cells to differentiate. During the progress period, we obtained substantial evidence for both of these hypotheses. With regard to the first hypothesis, we have used genome-wide approaches to identify hundreds of messenger RNAs that are regulated by NMD in both in vivo (in mice) and in vitro (in cell lines). To determine which of these messenger mRNAs are directly degraded by NMD, we have used a variety of approaches. This work has revealed that NMD preferentially degrades messenger RNAs encoding neural differentiation inhibitors and proliferation inhibitors in neural stem cells. In contrast, very few messenger RNAs encoding pro-stem cell proteins or pro-proliferation proteins are degraded by NMD. Together this provides support for our hypothesis that NMD promotes the stem-like state by shifting the proportion of messenger RNAs in a cell towards promoting an undifferentiated, proliferative cell state. During the progress period, we have obtained considerable evidence that this hypothesis not only applies to mouse stem cells but also human embryonic stem cells. With regard to the second hypothesis, we have found that many proteins that are directly involved in the NMD pathway are downregulated upon differentiation of stem and progenitor cells. Not only are NMD proteins reduced in level, but we find that the magnitude of NMD itself is reduced. We have used a variety of molecular techniques to determine whether this NMD downregulatory response has a role in neural differentiation and found that NMD downreglation is both necessary and sufficient for this event. Such experiments have also revealed particular messenger mRNAs degraded by NMD that are crucial for the NMD downregulatory response to promote neural differentiation. During the progress period, we obtained both experimental and genome-wide data that this applies to human embryonic stem cells. Our research has implications for intellectual disability cases in humans caused by mutations in an X-linked gene essential for NMD. Patients with mutations in this gene—UPF3B—not only have intellectual disability, but also schizophrenia, autism, and attention-deficit/hyperactivity disorder. Thus, the study of NMD may provide insight into a wide spectrum of cognitive and psychological disorders. We are currently in the process of generating and characterizing induced-pluripotent stem (iPS) cells from intellectual disability patients with mutations in the UPF3B gene towards this goal.
Funding Type: 
Basic Biology IV
Grant Number: 
RB4-06079
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$1 506 420
Disease Focus: 
Huntington's Disease
Neurological Disorders
Parkinson's Disease
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 

A major medical problem in CA is the growing population of individuals with neurodegenerative diseases, including Parkinson’s (PD) and Huntington’s (HD) disease. These diseases affect millions of people, sometimes during the prime of their lives, and lead to total incapacitation and ultimately death. No treatment blocks the progression of neurodegeneration. We propose to conduct fundamental studies to understand the basic common disease mechanisms of neurodegenerative disorders to begin to develop effective treatments for these diseases. Our work will target human stem cells made from cells from patients with HD and PD that are developed into the very cells that degenerate in these diseases, striatal neurons and dopamine neurons, respectively. We will use a highly integrated approach with innovative molecular analysis of gene networks that change the states of proteins in these diseases and state-of-the-art imaging technology to visualize living neurons in a culture dish to assess cause and effect relationships between biochemical changes in the cells and their gradual death. Importantly, we will test whether drugs effective in animal model systems are also effective in blocking the disease mechanisms in the human HD and PD neurons. These human preclinical studies could rapidly lead to clinical testing, since some of the drugs have already been examined extensively in humans in the past for treating other disorders and are safe.

Statement of Benefit to California: 

Neurodegenerative diseases, such as Parkinson’s (PD) and Huntington’s disease (HD), are devastating to patients and families and place a major financial burden on California. No treatments effectively block progression of any neurodegenerative disease. A forward-thinking team effort will allow highly experienced investigators in neurodegenerative disease and stem cell research to investigate common basic mechanisms that cause these diseases. Most important is the translational impact of our studies. We will use neurons and astrocytes derived from patient induced pluripotent stem cells to identify novel targets and discover disease-modifying drugs to block the degenerative process. These can be quickly transitioned to testing in preclinical and clinical trials to treat HD and other neurodegenerative diseases. We are building on an existing strong team of California-based investigators to complete the studies. Future benefits to California citizens include: 1) discovery and development of new HD treatments with application to other diseases, such as PD, that affect thousands of Californians, 2) transfer of new technologies and intellectual property to the public realm with resulting IP revenues to the state with possible creation of new biotechnology spin-off companies, and 3) reductions in extensive care-giving and medical costs. We anticipate the return to the State in terms of revenue, health benefits for its Citizens and job creation will be significant.

Progress Report: 
  • The goal of our study is to identify common mechanisms that cause the degeneration of neurons and lead to most neurodegenerative disorders. Our work focuses on the protein homeostasis pathways that are disrupted in many forms of neurodegeneration, including Huntington’s disease (HD) and Parkinson’s disease (PD). In this first reporting period we have made great progress in developing novel methods to probe the autophagy pathway in single cells. This pathway is involved in the turnover of misfolded proteins and dysfunction organelles. Using our novel autophagy assays, we have preliminary data that indicate that the autophagy pathway in neurons from HD patients is modulated compared to healthy controls. We have also begun validating small molecules that activate the autophagy pathway and we are now moving these inducers into human neurons from HD patients to see if they reduce toxicity or other disease related phenotypes. Using pathway analysis we have also identified specific genes within the proteostasis network that are modulated in HD. We are now testing whether modulating these genes in human neurons from HD patients can lead to a reduction in neurodegeneration. In the final part of this study we are investigating whether neurodegenerative diseases, such as HD and PD, share changes in similar genes or pathways, specifically those involved in protein homeostasis. We have now established a human neuron model for PD and have used it to identify potential targets that modulate the disease phenotype via changes in proteostasis. Using the assays, autophagy drugs and pathway analysis described above, we hope to identify overlapping targets that could potentially rescue disease associated phenotypes in both HD and PD.
  • The goal of our study is to identify common mechanisms that cause the degeneration of neurons and lead to most neurodegenerative disorders. Our work focuses on the protein homeostasis pathways that are disrupted in many forms of neurodegeneration, including Huntington’s disease (HD) and Parkinson’s disease (PD). In this reporting period we have made good progress in both developing new assays and novel autophagy compounds and identifying potential genetic targets that could lead to novel therapeutic strategies for patients with HD and PD. We have developed methods to measure the degradation rates of proteins involved in causing neurodegeneration and the decay rates of mitochondria that are disrupted during the progression of these diseases. We are now investigating if and how these degradation rates differ in cells from patients with HD. We have developed novel compounds that can activate the autophagy pathway which is critical for degrading the toxic proteins that cause neurodegeneration. We are now testing if these compounds can increase the survival of neurons derived from iPSCs from patients with HD. Using pathway analysis we have also identified specific genes within the proteostasis network that are modulated in HD. Specifically we have identified deubiquitinating enzymes as modulators of HD induced toxicity and autophagy modulation, potentially indicating that importance of the autophagy pathway in the disease progression. We are also using RNAseq analysis to investigate if neurons derived from iPSCs from PD patients exhibit differences in the genes expressed in the proteostasis network. If we identify key genes we will use our established human neuron model for PD to validate whether these genes modulate the disease phenotype via changes in proteostasis. Ultimately we hope to identify overlapping targets that could potentially rescue disease associated phenotypes in both HD and PD.
Funding Type: 
Basic Biology IV
Grant Number: 
RB4-06277
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$1 367 172
Disease Focus: 
Neurological Disorders
Pediatrics
oldStatus: 
Active
Public Abstract: 

Alexander disease (AxD) is a devastating childhood disease that affects neural development and causes mental retardation, seizures and spasticity. The most common form of AxD occurs during the first two years of life and AxD children show delayed mental and physical development, and die by the age of six. AxD occurs in diverse ethnic, racial, and geographic groups and there is no cure; the available treatment only temporally relieves symptoms, but not targets the cause of the disease. Previous studies have shown that specific nervous system cells called astrocytes are abnormal in AxD patients. Astrocytes support both nerve cell growth and function, so the defects in AxD astrocytes are thought to lead to the nervous system defects. We want to generate special cells, called induced pluripotent stem cells (iPSCs) from the skin or blood cells of AxD patients to create an unprecedented, new platform for the study and treatment of AxD. We can grow large quantities of iPSCs in the laboratory and then, using novel methods that we have already established, coax them to develop into AxD astrocytes. We will study these AxD astrocytes to find out how their defects cause the disease, and then use them to validate potential drug targets. In the future, these cells can also be used to screen for new drugs and to test novel treatments. In addition to benefiting AxD children, we expect that our approach and results will benefit the study of other, similar childhood nervous system diseases.

Statement of Benefit to California: 

It is estimated that California has approximately 12% of all US cases of AxD, a devastating childhood neurological disorder that leads to mental retardation and early death. At present, there is no cure or standard treatment available for AxD. Current treatment is symptomatic only. In addition to the tremendous emotional and physical pain that this disease inflicts on Californian families, it adds a medical and fiscal burden larger than that of any other states. Therefore, there is a real need to understand the underlying mechanisms of this disease in order to develop an effective treatment strategy. Stem cells provide great hope for the treatment of a variety of human diseases. Our proposal to establish a stem cell-based cellular model for AxD could lead to the development of new therapies that will represent great potential not only for Californian health care patients, but also for the Californian pharmaceutical and biotechnology industries. In addition to benefiting the treatment of AxD patients, we expect that our approach and results will benefit the study of other related neurological diseases that occur in California and the US.

Progress Report: 
  • Alexander disease (AxD) is a devastating childhood disease that affects neural development and causes mental retardation, seizures and spasticity. AxD children usually die by the age of six. AxD occurs in diverse ethnic, racial, and geographic groups and there is no cure; the available treatment only temporally relieves symptoms, but not targets the cause of the disease. Previous studies have shown that specific nervous system cells called astrocytes are abnormal in AxD patients. We generated special cells, called induced pluripotent stem cells (iPSCs) from the skin cells of AxD patients, and coaxed them to develop into AxD astrocytes. We will study these AxD astrocytes to find out how their defects cause the disease, and then use them to validate potential drug targets. In the future, these cells can also be used to screen for new drugs and to test novel treatments. In addition to benefiting AxD children, we expect that our approach and results will benefit the study of other, similar childhood nervous system diseases.
  • Alexander disease (AxD) is a devastating childhood disease that affects neural development and causes mental retardation, seizures and spasticity. AxD children usually die by the age of six. AxD occurs in diverse ethnic, racial, and geographic groups and there is no cure; the available treatment only temporally relieves symptoms, but not targets the cause of the disease. Previous studies have shown that specific nervous system cells called astrocytes are abnormal in AxD patients. We generated special cells, called induced pluripotent stem cells (iPSCs) from the skin cells of AxD patients, and coaxed them to develop into AxD astrocytes. We have been studying these AxD astrocytes to find out how their defects cause the disease and have identified a defective signaling pathway in these cells. In the future, these cells can also be used to screen for new drugs and to test novel treatments. In addition to benefiting AxD children, we expect that our approach and results will benefit the study of other, similar childhood nervous system diseases.
Funding Type: 
iPSC Consortia Award
Grant Number: 
RP1-05741
Investigator: 
Type: 
PI
Type: 
Partner-PI
ICOC Funds Committed: 
$300 000
Disease Focus: 
Huntington's Disease
Neurological Disorders
Collaborative Funder: 
NIH
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 
Statement of Benefit to California: 
Progress Report: 
  • Huntington’s disease (HD) is a significant neurodegenerative disease with unique genetic features. A CAG expansion in Huntington gene is correlated with severity and onset of sub-clinical and overt clinical symptoms, make it particularly suited to therapeutic development . The single genetic cause offers the opportunity to understand the pathological process triggered in all individuals with a CAG expansion, as emerging evidence suggests effects of the mutation in all cell types, though striatal neurons are most vulnerable to degeneration. Moreover, by virtue of a molecular test for the mutation, a unique opportunity exists to intervene/treat before the onset of overt clinical symptoms utilizing sub-clinical phenotypes emerging in pre-manifest individuals. Since human induced pluripotent stem cells (iPSCs) have the power to make any cell type in the human body, we are utilizing the technology to make patients iPSCs and study the effects of different number of CAG repeats on the neurons we generate from the patient iPS cells. Preliminary studies indicate that CAG length–dependent phenotypes occur at all stages of differentiation, from iPSC through to mature neurons and are likely to occur in non-neuronal cells as well, which can also be investigated using the iPSC that we are creating. The non-integrating technology (avoids integration of potentially deleterious reprogramming factors in the cell DNA) for producing iPSC lines is crucial to obtaining reproducible disease traits from patient cells.
  • The Cedars-Sinai RMI iPSC Core is part of the Huntington’s Disease (HD) consortium. In the past year the iPSC Core has made many new non-integrating induced pluripotent stem (iPSC) cell lines from HD patients with different numbers of CAG repeat expansions. The grant application proposed generation of 18 HD and Control iPSC lines. Instead we are generating 20 iPSC lines. So far we have already generated 17 iPSC lines from individuals with Huntington’s disease and controls (10 HD patients and 7 controls). In order to have the disease trait reproducible across multiple groups, three clonal iPSC lines were generated from each subject. Some of these lines have (or are in process) of expansion for distribution to consortium members. We are now in the process of making the last 3 lines as part of this grant application to generate a HD iPSC repository with total of 20 patient/control lines from subjects with multitude of CAG repeat numbers. Most of these lines have undergone rigorous battery of characterization for pluripotency determination, while some other lines are currently being validated through more characterization tests. Neural stem cell aggregates (EZ spheres) have been generated from few of the patient lines in the Svendsen lab (not supported by this grant). We have also submitted 6 patient iPSC lines to Coriell Cell Repository for larger banking and distribution of these important and resourceful lines to other academic investigators and industry. We strongly believe that this iPSC repository will enormously speed up the process of understanding the disease causing mechanisms in HD patient brain cells as well as discovering novel therapeutics or drugs that may one day be able to treat HD patients.
  • Huntington’s disease (HD) is a fatal neurodegenerative condition with no current treatment. This significant neurodegenerative disease, whose relatively simple and unique known genetic cause, a CAG expansion in the HD gene correlated with severity and onset of clinical symptoms, makes it particularly suited to therapeutic development. The Huntington’s disease (HD) iPS cell consortium, funded with NIH and CIRM support, brings together leading groups in stem cell and HD research to establish whether newly created iPS cell lines show HD related (i.e., CAG length-dependent) phenotypes. Human iPSC technology can be used to generate specific neuronal and glial cell types, permitting investigation of the effects of the genetic lesion in the susceptible human cell types in the context of HD. The monogenic nature of HD and the existence of allelic series of iPSCs with a range of CAG repeat lengths confer tremendous power to model HD. Through CIRM support this consortium has capitalized on new technologies to use non-integrating approaches for reprogramming and promising phenotypes in current HD iPS lines to develop robust and validated assays for drug development for HD.
  • Significant progress has been made through CIRM-funded support of this proposal. Notably, the Cedars-Sinai Medical Center’s Board of iPSC core housed in the Board of Governors Regenerative Medicine Institute has taken skin cells from HD patients with a wide range of CAG repeats (43 to 180), and unaffected healthy controls (21 to 33) and reprogrammed then to pluripotency using the latest non-integrating iPS cell technology. So far 18 well-characterized patient-specific iPSC lines have been generated. These new iPSC lines have been rigorously characterized by our iPSC core and available to HD research community throughout California and the world. The Svendsen lab and the other HD iPSC Consortium laboratories have already used these lines and differentiated into relevant neuronal cell types to study the disease mechanisms as well develop new treatment.
  • These cell lines will be an essential resource for academic groups and pharmaceutical companies for studying pathogenesis and for testing experimental therapeutics for HD. The ultimate goal is to develop and validate methods and assays using >96 well format for CAG repeat length-dependent phenotypes that are amenable to high content/throughput screening methods. Assays developed using these patient-specific iPSC lines and their neuronal derivatives will allow academic groups and pharmaceutical companies to study pathogenesis and test experimental therapeutics for HD, which will significantly advance both our understanding of HD and potential treatments for this devastating and currently untreatable disease.

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