Neurological Disorders

Coding Dimension ID: 
303
Coding Dimension path name: 
Neurological Disorders
Funding Type: 
Basic Biology V
Grant Number: 
RB5-07254
Investigator: 
Type: 
PI
ICOC Funds Committed: 
$1 003 590
Disease Focus: 
Neurological Disorders
Stem Cell Use: 
Adult Stem Cell
oldStatus: 
Closed
Public Abstract: 

Stem cells generate mature, functional cells after proteins on the cell surface interact with cues from the environment encountered during development or after transplantation. Thus, these cell surface proteins are critical for directing transplanted stem cells to form appropriate cells to treat injury or disease. A key modification regulating cell surface proteins is glycosylation, which is the addition of sugars onto proteins and has not been well studied in neural stem cells. We focus on a major unsolved problem in the neural stem cell field: do different proteins coated with sugars on the surfaces of cells in this lineage (neuron precursors, NPs and astrocyte precursors, APs) determine what types of mature cells will form? We hypothesize key players directing cellular decisions are glycosylated proteins controlling how precursors respond to extracellular cues. We will address this hypothesis with aims investigating whether (1) glycosylation pathways predicted to affect cell surface proteins differ between NPs and APs, (2) glycosylated proteins on the surface of NPs and APs serve as instructive cues governing fate or merely mark their fate potential, and (3) glycosylation pathways regulate cell surface proteins likely to affect fate choice. By answering these questions we will better understand the formation of NPs and APs, which will improve the use of these cells to treat brain and spinal cord diseases and injuries.

Statement of Benefit to California: 

The goal of this project is to determine how cell surface proteins differ between cells in the neural lineage that form two types of final, mature cells (neurons and astrocytes) in the brain and spinal cord. In the course of these studies, we will uncover specific properties of human stem cells that are used to treat neurological diseases and injuries. We expect this knowledge will improve the use of these cells in transplants by enabling more control over what type of mature cell will be formed from transplanted cells. Also, cells that specifically generate either neurons or astrocytes can be used for drug testing, which will help to predict the effects of compounds on cells in the human brain. We hope our research will greatly improve identification, isolation, and utility of specific types of human neural stem cells for treatment of human conditions. Furthermore, this project will generate new jobs for high-skilled workers and, hopefully, intellectual property that will contribute to the economic growth of California.

Progress Report: 
  • Overall, our biggest breakthrough this year has been the identification of a link among the sugars on the cell surface, a label free electrical measure reflecting the type of mature cell the stem cells will become (membrane capacitance), and stem cell fate potential, or the ability of the cell to form a particular type of mature cell. Stem cells generate mature, functional cells after proteins on the cell surface interact with cues from the environment encountered during development or after transplantation. Thus, these cell surface proteins are critical for directing transplanted stem cells to form the appropriate types of cells to treat injury or disease. A key modification regulating cell surface proteins is glycosylation, which is the addition of sugars onto proteins and has not been well studied in neural stem cells. Our project focuses on a major unsolved problem in the neural stem cell field: do different proteins coated with sugars on the surfaces of cells in this lineage (neuron precursors, NPs and astrocyte precursors, APs) determine what types of mature cells will form? We hypothesize key players directing cellular decisions are glycosylated proteins controlling how precursors respond to extracellular cues. This year on the project, we found a particular glycosylation pathway that adds highly branched sugars regulates cell surface properties and controls the decision to form either a neuron or an astrocyte. In the next year of the project, we will explore this pathway further and perform experiments to identify the proteins on the cell surface important for determining the formation of either mature neurons or astrocytes. By answering these questions, we will better understand the regulation of NPs and APs, which will improve the use of these cells to treat brain and spinal cord diseases and injuries.
Funding Type: 
Basic Biology V
Grant Number: 
RB5-07363
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$1 178 370
Disease Focus: 
Stroke
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Closed
Public Abstract: 

Stem cell therapy holds promise for the almost million Americans yearly who suffer a stroke. Preclinical data have shown that human neural stem cells (hNSCs) aid recovery after stroke, resulting in a major effort to advance stem cell therapy to the clinic, and we are currently transitioning our hNSC product to the clinic for stroke therapy. In this proposal we will explore how these cells improve lost function. We have already shown that injected hNSCs secrete factors that promote the gross rewiring of the brain, a major component of the spontaneous recovery observed after stroke. We now intend to focus on the connections between neurons, the synapses, which are a critical part of this rewiring process. We aim to quantify the effect of hNSCs on synapse density and function, and explore whether the stem cells secrete restorative synaptogenic factors or form functional synapses with pre-existing neurons. Our pursuit is made possible by our combination of state-of-the-art imaging techniques enabling us to visualize, characterize, and quantify these tiny synaptic structures and their interaction with the hNSCs. Furthermore, by engineering the hNSCs we can identify the factors they secrete in the brain and identify those which modulate synaptic connections. Our proposed studies will provide important insight into how transplanted stem cells induce recovery after stroke, with potential applicability to other brain diseases.

Statement of Benefit to California: 

Cerebrovascular stroke is the fourth leading cause of mortality in the United States and a significant source of long-term physical and cognitive disability that has devastating consequences to patients and their families. In California alone, over 9% of adults 65 years or older have had a stroke according to a 2005 study. In the next 20 years the societal toll is projected to amount to millions of patients and 18.8 billion dollars per year in direct medical costs. To date, there is no approved therapeutic agent for the recovery phase after stroke, making the long-term care of stroke patients a tremendous socioeconomic burden that will continue to rise as our aging population increases. Our laboratory and others have demonstrated the promise of stem cell transplantation to treat stroke. We are dedicated to developing human neural stem cells (hNSCs) as a novel neuro-restorative treatment for lost motor function after stroke. The goal of our proposed work is to further understand how transplanted hNSCs improve stroke recovery, as dissecting the mechanism of action of stem cells in the stroke brain will ultimately improve the chance of clinical success. This could potentially provide significant cost savings to California, but more importantly benefit the thousands of Californians and their families who struggle with the aftermath of stroke.

Funding Type: 
Basic Biology V
Grant Number: 
RB5-06935
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$1 174 943
Disease Focus: 
Parkinson's Disease
Neurological Disorders
Stem Cell Use: 
iPS Cell
oldStatus: 
Closed
Public Abstract: 

Parkinson’s disease (PD), is one of the leading causes of disabilities and death and afflicting millions of people worldwide. Effective treatments are desperately needed but the underlying molecular and cellular mechanisms of Parkinson’s destructive path are poorly understood. Mitochondria are cell’s power plants that provide almost all the energy a cell needs. When these cellular power plants are damaged by stressful factors present in aging neurons, they release toxins (reactive oxygen species) to the rest of the neuron that can cause neuronal cell death (neurodegeneration). Healthy cells have an elegant mitochondrial quality control system to clear dysfunctional mitochondria and prevent their resultant devastation. Based on my work that Parkinson’s associated proteins PINK1 and Parkin control mitochondrial transport that might be essential for damaged mitochondrial clearance, I hypothesize that in Parkinson’s mutant neurons mitochondrial quality control is impaired thereby leading to neurodegeneration. I will test this hypothesis in iPSC (inducible pluripotent stem cells) from Parkinson’s patients. This work will be a major step forward in understanding the cellular dysfunctions underlying Parkinson’s etiology, and promise hopes to battle against this overwhelming health danger to our aging population.

Statement of Benefit to California: 

Parkinson's disease (PD), one of the most common neurodegenerative diseases, afflicts millions of people worldwide with tremendous global economic and societal burdens. About 500,000 people are currently living with PD in the U.S, and approximate 1/10 of them live in California. The number continues to soar as our population continues to age. An effective treatment is desperately needed but the underlying molecular and cellular mechanisms of PD’s destructive path remain poorly understood. This proposal aims to explore an innovative and critical cellular mechanism that controls mitochondrial transport and clearance via mitophagy in PD pathogenesis with elegant employment of bold and creative approaches to live image mitochondria in iPSC (inducible pluripotent stem cells)-derived dopaminergic neurons from Parkinson’s patients. This study is closely relevant to public health of the state of California and will greatly benefit its citizens, as it will illuminate the pathological causes of PD and provide novel targets for therapuetic intervention.

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: 
Early Translational IV
Grant Number: 
TR4-06747
Investigator: 
Type: 
Partner-PI
ICOC Funds Committed: 
$1 824 719
Disease Focus: 
Autism
Neurological Disorders
Rett's Syndrome
Pediatrics
Collaborative Funder: 
NIH
Stem Cell Use: 
iPS Cell
oldStatus: 
Active
Public Abstract: 

Autism spectrum disorders (ASD) are complex neurodevelopmental diseases that affect about 1% of children in the United States. Such diseases are mainly characterized by deficits in verbal communication, impaired social interaction, and limited and repetitive interests and behavior. The causes and best treatments remain uncertain. One of the major impediments to ASD research is the lack of relevant human disease models. Reprogramming of somatic cells to a pluripotent state (induced pluripotent stem cells, iPSCs) has been accomplished using human cells. Isogenic pluripotent cells are attractive from the prospective to understanding complex diseases, such as ASD. The main goal of this project is to accelerate drug discovery to treat ASD using astrocytes generated from human iPSC. The model recapitulates early stages of ASD and represents a promising cellular tool for drug screening, diagnosis and personalized treatment. By testing whether drugs have differential effects in iPSC-derived astrocytes, we can begin to unravel how genetic variation in ASD dictates responses to different drugs. Insights that emerge from our studies may drive the development of new therapeutic interventions for ASD. They may also illuminate possible differences in drug responsiveness in different patients and potentially define a molecular signature resulting from ASD variants, which could predict the onset of disease before symptoms are seen.

Statement of Benefit to California: 

Autism spectrum disorders, including Rett syndrome, Angelman syndrome, Timothy syndrome, Fragile X syndrome, Tuberous sclerosis, Asperger syndrome or childhood disintegrative disorder, affect many Californian children. In the absence of a functionally effective cure or early diagnostic tool, the cost of caring for patients with such pediatric diseases is high, in addition to a major personal and family impact since childhood. The strikingly high prevalence of ASD, dramatically increasing over the past years, has led to the emotional view that ASD can be traced to a single source, such as vaccine, preservatives or other environmental factors. Such perspective has a negative impact on science and society in general. Our major goal is to develop a drug-screening platform to rescue deficiencies showed from brain cells derived from induced pluripotent stem cells generated from patients with ASD. If successful, our model will bring novel insights on the dentification of potential diagnostics for early detection of ASD risk, or ability to predict severity of particular symptoms. In addition, the development of this type of pharmacological therapeutic approach in California will serve as an important proof of principle and stimulate the formation of businesses that seek to develop these types of therapies (providing banks of inducible pluripotent stem cells) in California with consequent economic benefit.

Progress Report: 
  • The progress in our research regarding the role of human astrocytes in Rett syndrome (RTT) showed us that RTT-derived astrocyte display several phenotypes that illustrate its differences compared to healthy control astrocytes (WT). RTT astrocytes are unable to propagate calcium wave when mechanically stimulated . In addition to that, when placed in medium that contains glutamate, the natural uptake and buffering of this compound is impaired in RTT-derived astrocytes. Furthermore, when WT neurons are placed on top of RTT astrocytes, there is a clear the negative effect of these cells in neuronal homeostasis. Remarkably, WT astrocytes are able to rescue RTT neuronal phenotypes when in direct contact, illustrating the important role that astrocytes have in maintaining neuronal viability and maturation. Several mis-regulation in gene expression pathways indicated those phenotypes, both in calcium and glutamate dependent genes. Strikingly, further genetic analysis led us to identify several mis-regulations in pro-inflammatory cytokines. Multiplex ELISA platforms also pointed towards a difference in cytokine secretion between WT and RTT syndrome astrocytes, being the RTT cells illustrative of a pro-inflammatory scenario. We have define one of these secreted cytokines as our primary read out for the HT-screening. We are now facing a transportation issue with very sensitive cells, but have an innovative plan to make it to work and also get some quick results that may have clinical relevance.
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: 
New Faculty Physician Scientist
Grant Number: 
RN3-06530
Investigator: 
Type: 
PI
ICOC Funds Committed: 
$3 031 737
Disease Focus: 
Neuropathy
Neurological Disorders
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Active
Public Abstract: 

The applicant is an MD/PhD trained physician scientist, whose clinical expertise is neuromuscular disorders including peripheral nerve disease. The proposal is aimed at providing a research proposal and career development plan that will allow the applicant to develop an independent research program, which attempts to bring stem cell based therapies to patients with peripheral nerve diseases. The proposal will use “adult stem cells” derived from patients with an inherited nerve disease, correct the genetic abnormality in those cells, and determine the feasibility of transplanting the genetically engineered cells back into peripheral nerve to slow disease progression.

Statement of Benefit to California: 

The proposed research will benefit the State of California as it will support the career development of a uniquely trained physician scientist to establish an innovative translational stem cell research program aimed toward direct clinical application to patients. The cutting edge technologies proposed are directly in line with the fundamental purpose of the California Initiative for Regenerative Medicine. If successful, both scientific and patient advocate organizations would recognize that these advances came directly from the unique efforts of CIRM and the State of California to lead the world in stem cell research. Finally, as a result of funding of this award, further financial investments from private and public funding organizations would directly benefit the State in the years to come.

Progress Report: 
  • During this award period we have made significant progress. We have established induced pluripotent stem cell (iPSC) lines from four patients with Charcot-Marie-Tooth disease type 1A (CMT1A) due to the PMP22 duplication. We have validated our strategy to genetically engineer induced pluripotent stem cells from patients with inherited neuropathy, and have genetically engineered several patient lines. We further have begun to differentiate these iPSCs into Schwann cell precursors, to begin to investigate cell type specific defects that cause peripheral neurodegeneration in patients with CMT1A. Finally we have imported and characterized a transgenic rat model of CMT1A in order to begin to investigate the ability to inject iPSC derived Schwann cell precursors into rodent nerves as a possible neuroprotective strategy.
  • During this reporting period we developed genetically corrected induced pluripotent stem cell lines from patients with CMT1A. We improved and validated a novel method for differentiating Schwann cells from iPSCs, and used this to generate human Schwann cells from patients and controls. Finally we have initiated pilot studies injecting human iPSC derived Schwann cells into the peripheral nerves of rats with myelin diseases to determine whether cell replacement therapy is a viable strategy in these disorders.
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.

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