Diabetes

Coding Dimension ID: 
289
Coding Dimension path name: 
Diabetes
Funding Type: 
Alpha Stem Cell Clinics
Grant Number: 
AC1-07764
Investigator: 
ICOC Funds Committed: 
$8 000 000
Disease Focus: 
Diabetes
Spinal Cord Injury
Neurological Disorders
Stem Cell Use: 
Embryonic Stem Cell
Adult Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
Adult Stem Cell
Public Abstract: 

The proposed alpha clinic will bring together an outstanding team of physician-scientists with substantial clinical trials experience including stem cell and other cellular treatments of blood diseases and others. This team will also draw on our unique regional competitive advantages derived from our history of extensive collaboration with investigators at many nearby first-class research institutions and biotech companies. We propose to include these regional assets in our plans to translate our successful research on basic properties of stem cells to stem cell clinical trials and ultimately to delivery of effective and novel therapies. We propose to build an alpha clinic that serves the stem cell clinical trial needs of our large region where we are the only major academic health center with the needed expertise to establish a high impact alpha clinic. Our infrastructure will initially be developed and then used to support two major high-impact stem cell clinical trials: one in type I diabetes and one in spinal cord injury. Both are collaborations with established and well known companies. The type I diabetes trial will test embryonic stem cell derived cells that differentiate to become the missing beta cells of the pancreas. The cells are contained in a semipermeable bag that has inherent safety because of restriction of cell migration while allowing proper control of insulin levels in response to blood sugar. These hybrid devices are implanted just beneath the skin in patients in these trials. In a second trial of stem cell therapy for spinal cord injury, neuronal stem cells that have been shown to have substantial safety and efficacy in animal models of spinal cord injury and other types of spinal cord trauma or disease will be tested in human patients with chronic spinal cord injury. Both of these trials have the potential to have very substantial and important impact on patients with these diseases and the families and society that supports them. Following on these two trials, we are planning stem cell clinical trials for heart failure, cancer, ALS, and other terrible deadly disorders. Our proposed alpha clinic also benefits from very substantial leveraged institutional commitments, which will allow for an alpha clinic that is sustainable well beyond the five-year grant, which is essential to continue to manage the patients who have participated in the first trials being planned since multi-year followup and tracking is essential scientifically and ethically. We have a plan for our proposed alpha clinic to be sustainable to 10 years and beyond to the point at which these therapies if successful will be delivered to patients in our healthcare system.

Statement of Benefit to California: 

Many terrible diseases that afflict the citizens of California and cause substantial economic and emotional disruption to California families can potentially be treated with novel stem cell therapies. These therapies need to be tested in a rigorous and unbiased fashion in clinical trials, which is the focus of our proposed alpha clinic. Our clinic proposes to begin with clinical trials in two major diseases in need of improved treatment: type I diabetes and spinal cord injuries. The type I diabetes clinical trial will test a novel hybrid embryonic stem cell-derived pancreatic cell/encapsulation technology that is implanted just beneath the skin in an out-patient procedure, and is inherently safe because the cells are confined to a semi-permeable bag. The spinal cord injury trial will test the benefit of neural stem cells delivered to the site of injury. Both have substantial positive evidence in animal models and have the potential of leading to major breakthroughs. In addition to providing the infrastructure for these two trials, our proposed alpha clinic will also take advantage of very substantial regional expertise at our partner institutions to test stem cells in other diseases of importance in California including heart failure, ALS, cancer, and many others. Our proposed alpha clinic will also be a major economic as well as medical driver as it leverages substantial institutional and private sector commitment, and has the potential to deliver breakthrough therapies that will be marketed either in a health care system or by private sector companies.

Funding Type: 
Accelerated Development Pathway I
Grant Number: 
AP1-08039
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$16 603 160
Disease Focus: 
Diabetes
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
Public Abstract: 

We are developing a stem cell-derived replacement cell therapy for insulin-requiring diabetes. Through a process known as directed differentiation, embryonic stem cells are turned into pancreatic cells in the laboratory. The pancreatic cells are loaded into a delivery device, which is essentially a small envelope made with a semi-permeable membrane, not unlike a flat tea bag. When the cells in the device (combination product) are implanted under the skin, they become pancreatic endocrine cells, including insulin-producing beta cells that respond to elevated blood glucose by releasing insulin in a physiologic manner. The prototype combination product has been tested in hundreds of animals, is routinely curative in a mouse model of chemically-induced diabetes, and has been shown to be safe in several animal studies. Moreover, the delivery device has been shown to protect cells from a recipient’s immune system. The Team has received valuable feedback from the FDA, and we plan to launch the first clinical test of our therapeutic candidate in patients with diabetes in 2014. This first clinical trial will utilize the prototype to establish safety in humans, and determine the dosing range that might provide benefit to patients with diabetes.

The current application is to fund additional clinical research, and associated product development activity, that will (1) ensure the first trial is executed in a most informative and timely fashion, (2) accelerate the pace at which information is collected on how the product works in humans – testing various formats, and in different types of patients – and (3) substantially increase the likelihood that the most appropriate format and patient population is selected for a definitive “Phase 3” clinical trial. A Phase 3 trial serves as the basis for an application to the FDA to obtain a license to market the product. In this way, CIRM Accelerated Development Pathway designation of the project will substantially increase the probability that, and pace at which, this product concept becomes a real treatment available to the millions of patients in need.

Statement of Benefit to California: 

Diabetes mellitus currently afflicts approximately 370 million people worldwide, with projections of over 550 million by the year 2030 (sources: World Health Organization; International Diabetes Federation). In the year 2000 there were approximately 2 million cases of diabetes in California (source: Diabetes Control Program, California Department of Health Services). Further, the disease disproportionately affects certain minority groups and the elderly. Despite the use of insulin and advances in its delivery, the human cost of diabetes is underscored by the financial costs to society: tens of billions of dollars each year in California alone. The primary cause of type 1 diabetes, and contributing significantly to type 2 diabetes as well, is the loss of insulin-producing pancreatic beta cells. The CIRM Diabetes Disease Team Project is developing an innovative beta cell replacement therapy for insulin-requiring diabetes. If successful, the therapy will go beyond insulin function, and will perform the full array of normal beta cell functions, including responding in a more physiological manner than manual or mechanized insulin self-administration. Because they will be more physiological, the replacement cells could reduce the long-term effects of diabetes. Moreover, the cell therapy will alleviate patients of the constant monitoring of blood glucose, painful insulin injections, and the ever-present risk of overdosing with insulin. For these reasons, it is possible that the product could transform the diabetes treatment landscape dramatically and even replace pharmaceutical insulin in the market. This product will be available in California first, through clinical testing, and if approved by the FDA for commercial production, will eventually help hundreds of thousands of Californians with diabetes. The product will substantially increase quality of life for patients and their families, while significantly reducing the health care burden in the state. The proposed project will employ Californian doctors and scientists, and success will prove highly noteworthy for the state. Lastly, once commercially marketed, the product will yield additional jobs in manufacturing, sales, and related industries, and generate revenue for California. Given the market need and the clear feasibility, the product could become the most significant stem cell-based medical treatment of the coming decade, and that will be a tremendous achievement for California, its taxpayers, and CIRM.

Funding Type: 
Strategic Partnership I
Grant Number: 
SP1-06513
Investigator: 
Institution: 
Type: 
PI
ICOC Funds Committed: 
$10 075 070
Disease Focus: 
Diabetes
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 

Diabetes exacts a tremendous toll on patients, their families, and society. Autoimmune Type 1 diabetes, often called juvenile-onset diabetes, is caused by a person’s own immune system mistakenly destroying their insulin-producing cells in the pancreas, known as beta cells. When those beta cells are lost, the ability to produce insulin in response to consumed carbohydrates is lost, and blood sugar can increase to toxic levels. Although not due to autoimmunity, Type 2 diabetics often lose their ability to produce insulin as well. While pharmaceutical insulin is commonly used to control both types of diabetes, it is difficult to self-administer optimally, does not sufficiently replace beta cells, and the adverse short- and long-term effects of diabetes and risks associated with insulin usage remain, including potentially fatal hypoglycemic episodes, nerve damage, blindness, kidney failure, foot ulcers / amputations, and heart disease.

Ideally, one would like to replace lost beta cells, and attempts to do so have included the use of pancreas transplants, beta cell (islet) transplants, and transplants of animal cells. Unfortunately, those approaches are hindered by 1) a limited amount of donor tissue, and 2) issues regarding immunological incompatibility between donors and recipients. To solve the first problem, the group applying for this CIRM award has developed methods to make replacement beta cells from human embryonic stem cells (hESC), which can be reliably grown in large-scale batches. The hESC-derived beta cells have been shown to cure experimental diabetes in mice and rats. Regarding the issue of donor-recipient compatibility, the group has found that the cells can be administered under the skin in a simple device, essentially an envelope made of semi-permeable membrane, which is intended to protect the implanted cells from the patient’s immune system. Upon implant, the cell-loaded device, which also keeps the implanted cells in place, acquires its own dedicated circulation. This blood supply provides oxygen and nutrients to the implanted cells, and also allows them to respond to blood sugar by releasing pancreatic hormones such as insulin into the circulation. Thus, the implanted cell-loaded device in essence represents a “replacement endocrine pancreas” with its own protection from autoimmunity. This product could return a patient's blood sugar regulation to normal and alleviate both the day-to-day and long-term issues of diabetes.

The group has made tremendous progress in moving the product from concept through years of research and development. At this point an array of detailed work on the exact format to be tested in humans needs to be completed and submitted to the FDA on the way to clinical trials. The proposed award would provide critical funding, including potentially triggering matching funding from a large corporate partner, to advance the product through the first-in-human testing which will be very informative.

Statement of Benefit to California: 

Diabetes mellitus currently afflicts approximately 350 million people worldwide, with projections of over 500 million by the year 2030 (sources: World Health Organization; International Diabetes Federation). In the year 2000 there were an estimated 2,089,657 cases of diabetes in California (diagnosed + undiagnosed; source: Diabetes Control Program, California Department of Health Services). Further, the disease disproportionately affects certain minority groups and the elderly. Despite the use of insulin and advances in its delivery, the human cost of diabetes is underscored by the financial costs to society: tens of billions of dollars each year in California alone. The primary cause of Type 1 diabetes, and contributing significantly to Type 2 diabetes as well, is the loss of insulin-producing pancreatic beta cells. The proposed Partnership will develop a beta cell replacement therapy for insulin-dependent diabetes. If successful, the therapy will go beyond insulin function, and will perform the full array of normal beta cell functions, including responding in a more physiological manner than manual or mechanized insulin administration. Because they will be more physiological, the replacement cells should also reduce the long-term effects of diabetes. Moreover, the cell therapy will alleviate patients of the constant monitoring of blood glucose and painful insulin injections. For these reasons, it is possible that the product could transform the diabetes treatment landscape dramatically and even replace pharmaceutical insulin in the market. This product will be available in California first, through clinical testing, and if approved by the FDA for commercial production, will eventually help hundreds of thousands of Californians with diabetes. The product will substantially increase quality of life for patients and their families while significantly reducing the health care burden in the state. The proposed Partnership will employ Californian doctors and scientists, and success will generate accolades and notoriety for the state. Lastly, once commercially marketed, the product will yield additional jobs in manufacturing, sales, and related industries, and generate revenue for California. Given the market need and the clear feasibility, the product could become the most significant stem cell-based medical treatment of the coming decade, and that will be a great achievement for California, its taxpayers, and CIRM.

Progress Report: 
  • In type 1 diabetes, the insulin-producing beta cells of the endocrine pancreas are destroyed by a person’s own immune system in a disease process known as autoimmunity. Consequently, those with type 1 diabetes are unable to produce insulin, which is necessary for control of blood glucose, and ultimately, for life. While pharmaceutical insulin provides life-saving relief, numerous risks and complications exist for those contending with this disease. A large body of research indicates that replacing the lost beta cells of the endocrine pancreas would represent a substantially more effective approach to treating type 1 diabetes.
  • To address this need, ViaCyte is developing a cell replacement therapy for type 1 diabetes. The proposed product is comprised of human pancreatic progenitor cells, known as PEC-01™ cells, loaded into and implanted within a flat encapsulation device or packet, known as the Encaptra® drug delivery system. PEC-01 cells are manufactured in the laboratory from human embryonic stem cells using a proprietary process that was developed at ViaCyte. The Encaptra device is made of biocompatible materials and is designed to retain PEC-01 cells in a specific location in the body, and protect them from a patient’s immune system. Together, PEC-01 cells within the Encaptra device are known as the VC-01™ combination product candidate, and animal studies have shown that following implant, the cells mature into functional human pancreatic endocrine cells (including insulin producing cells) that respond to changes in blood glucose appropriately, including the release of insulin into the circulation when blood glucose levels are increased.
  • The Strategic Partnership Award provides funding to support the preparation and launch of clinical testing of the VC-01 product candidate. The first year of the award was to support pre-clinical work including the preparation and filing of regulatory documents including, importantly, an Investigational New Drug (IND) application with the FDA, to allow testing in human subjects to commence.
  • The team made great progress in the first year of the award and achieved all planned milestones in a timely fashion. Specifically, the group was able to complete the necessary animal safety and efficacy studies (performed under Good Laboratory Practices) that are prerequisite to human testing, obtain a license from the California Food and Drug Branch to allow product manufacturing to proceed, and manufacture PEC-01 cells (under Good Manufacturing Practices) and Encaptra devices (under Quality Systems Regulations) for the Phase 1/2 first-in-human clinical trial. Moreover, the group established and finalized materials and methods to load PEC-01 cells into Encaptra devices and prepare the sealed VC-01 combination product for transport to the point of service. The group also established storage conditions for the VC-01 product consistent with the logistics of the clinical trial. Lastly, the clinical plan and protocol were completed and the IND was submitted. Within 30 days the IND was allowed by the FDA, and the first clinical site was prepared to launch the trial.
Funding Type: 
Basic Biology V
Grant Number: 
RB5-07398
Investigator: 
Type: 
PI
ICOC Funds Committed: 
$526 896
Disease Focus: 
Diabetes
Cell Line Generation: 
Adult Stem Cell
oldStatus: 
Closed
Public Abstract: 

Patients with end-stage type 1 diabetes (T1D) can be effectively managed by allogeneic islet transplantation. However, a severe cadaveric organ shortage greatly limits use of this promising procedure. Stem cells have the potential to provide a solution to this bottleneck because of their ability to self-renew and differentiate into islet β-cells. Although progress has been made in coaxing human embryonic stem (ES) cells to differentiate into pancreatic progenitor-like cells in culture, there are safety concerns regarding ES cell-derived products because of their ability to form teratomas in vivo. In contrast, adult tissue cells lack teratoma potential. Our goal is to develop, for transplantation, insulin-expressing cells derived from adult human pancreatic progenitor-like cells. If successful, the proposed research will establish a new paradigm for the development of cell products derived from adult pancreata and enable important advances in cell replacement therapy for T1D. This research will allow human cadaveric adult pancreatic tissues, which are largely discarded after islet isolation, to be used to maximum efficiency in transplantation. Moreover, the results of these studies will be applicable to the treatment of end-stage type 2 diabetes patients, in whom islet β-cells are exhausted and dysfunctional.

Statement of Benefit to California: 

In type 1 and some type 2 diabetic patients, the pancreatic β-cells, which secrete insulin in response to elevated glucose concentrations in the blood, are insufficient or dysfunctional. Insulin injection is the most common form of therapy to control diabetes. However, insulin injection cannot match the physiological response conferred by endogenous β-cells, and complications inevitably develop over time. Allogeneic islet transplantation is beneficial to those diabetic patients who have developed end-stage complications. However, it is estimated that fewer than 1% of Californians most in need of islet transplantation can benefit from the procedure because there is a severe shortage of human cadaveric pancreas organs. This dire situation has led to the search for alternative sources of β-cells for transplantation. If human adult pancreatic stem and progenitor cells can be coaxed to differentiate into β-like cells in culture, they would provide large numbers of cells for replacement therapy. This proposal addresses the important challenge of producing β-cells through differentiation of human pancreatic stem and progenitor cells, with the ultimate objective of developing new treatments for diabetic patients.

Progress Report: 
  • This award has supported the development of several new classes of artificial proteins designed to facilitate production of glucose-sensitive cells for the treatment of Type 1 diabetes. We are exploring methods for controlled differentiation of glucose-sensitive cells from progenitor cells isolated from adult human pancreas. In our first year, we have prepared two new classes of artificial extracellular matrix (aECM) proteins, achieved targeted levels of elasticity in aECM gels to be used for cell encapsulation, demonstrated that cells can be encapsulated in and released from aECM gels without loss of viability, and shown that encapsulated cells maintain their insulin expression levels for at least 24 hours. These experiments lay the groundwork for the development of an important new source of cells for treatment of diabetic patients.
Funding Type: 
Basic Biology V
Grant Number: 
RB5-07262
Investigator: 
ICOC Funds Committed: 
$1 191 000
Disease Focus: 
Immune Disease
HIV/AIDS
Diabetes
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
oldStatus: 
Closed
Public Abstract: 

Stem cell research offers the promise of replacing missing or damaged tissues in the treatment of disease. Stem-cell-derived transplants still face problems with rejection as in traditional organ transplants. Several drugs can prevent rejection but also suppress the immune system, leaving patients vulnerable to infections and cancer. To avoid rejection without using drugs requires re-educating the immune system to “tolerate” the transplant and not see it as foreign. Because of its role in educating developing immune cells, the thymus is a critical organ in establishing what the immune system recognizes as “self” and not foreign, in a process known as immune tolerance. By growing a new thymus from stem cells matched to transplanted tissues, we can condition the immune system to be tolerant to the transplant and avoid chronic immunosuppression. We have developed a method to grow stem cells into thymic cells that become normal thymus tissue when grafted into mouse models. Notably, the new thymus can promote normal development of immune cells, indicating the potential for generating new, tolerant immune cells. We propose to induce immune tolerance to other stem-cell derived tissues using stem-cell-derived thymus tissue to engineer tolerance. We will optimize our methods of growing thymus tissue, which will be used to condition mice to accept stem-cell-derived pancreas grafts, testing their ability both to prevent rejection and to cure diabetes in a transplant model.

Statement of Benefit to California: 

The proposed work aims to improve the effectiveness of stem cell treatments by preventing immunological rejection of transplanted tissue derived from stem cells. An important barrier to the clinical use of stem-cell-derived organs and tissues is the potential of the immune system to reject or damage this regenerated tissue. Improved approaches to address immune rejection are needed since stem cell therapies are underway in treating diseases that have a wide impact on the health of Californians, including diabetes, Parkinson’s disease, Alzheimer’s disease, retinal eye diseases, and musculoskeletal diseases.

The proposed studies will improve treatment for these diseases by providing a novel method to halt immunologic rejection or destruction of tissues that are derived from stem cells. We have successfully developed methods to grow thymus tissue, which controls the ability of the immune system to be “tolerant” of transplanted tissue. Here we will improve methods to generate thymus from stem cells and show that it can promote survival of transplanted tissue derived from the same cells. By using the thymus to condition the immune system towards tolerance, we hope to avoid immune rejection without the use of immunosuppressive drugs. Induction of a tolerant immune system in this way would represent a significant advance in improving stem cell therapies. Thus, this work could have a broad impact on a large number of the disease treatments that involve stem cells.

Progress Report: 
  • We continue to make progress with our efforts to generate functional thymic epithelial cells that are derived from stem cell sources. Over the last year we have been able to improve our ability to differentiate thymic epithelial progenitors by using a 3-Dimenstional culture system. This system has improved our efficiency and we are currently further refining it for use in our differentiation method. In future years, this will help accelerate our progress in TEP generation. A second area that we have made progress in is the generation of a reporter stem cell line for a key transcription factor called FOXN1. These cells express a key marker that help tell us how well our protocol is working. Through the use of this new cell line we again have made substantial progress in our differentiation efficiency. In looking forward to the next years of funding, we are well positioned for our more elaborate experiments for looking at immune tolerance that is induced by our cells.
Funding Type: 
Preclinical Development Awards
Grant Number: 
PC1-08118
Investigator: 
Type: 
PI
Name: 
Type: 
Co-PI
ICOC Funds Committed: 
$5 039 008
Disease Focus: 
Diabetes
Skin Disease
Stem Cell Use: 
Adult Stem Cell
Public Abstract: 

The goal of our CIRM-funded Early Translational (ETA) grant was to engineer a product to improve healing in diabetic foot ulcers, a devastating consequence of diabetes that occurs in about 25% of all diabetic patients and is responsible for most leg or foot amputations. More than 6 million people in the US and up to 91 million people worldwide have diabetic foot ulcers (DFU). There is a clear medical need. There are products on the market that can improve wound healing for some, but not all patients. This causes a large financial burden for the health care system, and great suffering for the patients who live with open wounds, often infected, that progress to amputations. Therefore there is a clear medical need for advanced therapies to heal diabetic ulcers faster.

We proposed to create a combination product consisting of a scaffold for dermal regeneration (SDR) populated with human allogeneic mesenchymal stem cells (MSC) that have been pre-conditioned for optimized reparative function. We formed a team of established wound and stem cell/matrix experts, and this team has indeed successfully engineered and demonstrated efficacy of the preconditioned MSC-SDR in two animal models, and is now ready to progress to further dose-finding and initial biosafety studies in support of our very promising Development Candidate.

During the Early Translational grant, we developed a product that consists of an FDA-approved scaffold for dermal regeneration (SDR) filled with human bone marrow-derived Mesenchymal Stem Cells (MSC). These are then pre-incubated for 2 days in hypoxia and in the presence of a beta adrenergic antagonist. We have completed studies that demonstrate that this “next generation” stem cell product is highly efficacious in healing diabetic skin wounds, using mouse skin wound models in diabetic mice that have impaired and delayed healing and a porcine model.

In the PreClinical Development award period we propose to bring this product closer to clinical use for human patients. We propose dose finding studies to achieve the optimal dose with the largest safety margin. We will use a large animal wound model where skin wounds more closely resemble those in humans, to carry out these efficacy and early safety studies. We will use this time to create a Master Cell Bank of pure and effective human MSCs and to generate standard operating procedures to move us into the clinical arena. Finally, we will prepare a package for presentation to the FDA for moving the preclinical product forward toward a Phase 1/II clinical trial that will demonstrate efficacy and safety of the product in affected patients.

Statement of Benefit to California: 

While the number of individuals with all forms of chronic wounds is increasing in the general population, particularly with the rise of diabetes and aging of the population, the number of individuals affected by diabetic foot ulcers (DFU), the target disease for the development candidate in this proposal, is increasing in California at an alarming rate. That is because the prevalence of type 2 diabetes is now increasing within the state of California to epidemic proportions. In 2002, over one million California adults age 45 and older were diagnosed with diabetes, and by 2011 that number had risen to 2.3 million: 8.4% of the California population (1).

For reasons that are not all that clear, there are marked differences in the prevalence of diabetes in different Californian ethnic and racial groups. Among Californians 65 and older, diabetes is significantly more common in African Americans (25.6%) , and Latinos ( 24.4%) as compared to Caucasians (12.2%). (1) The diabetes brings with it devastating health impacts: it is the sixth most common cause of death in the United States. Among the morbidities associated with diabetes, DFU is one of the most debilitating. Approximately 15-25 percent of patients with diabetes will develop DFU, and of those, six percent will be hospitalized due to infection or other ulcer-related complications. According to a recent census, DFU is the leading cause of lower limb amputation and greater than 85% of amputations are preceded by an active foot ulcer.

Sadly for our state, we lead others in the US in the prevalence of DFU: "Of the 45 areas (44 states and DC) that reported information to the Behavioral Risk Factor Surveillance System, the world's largest, on-going telephone health survey system, the BRFSS diabetes module shows that Indiana (16.3%), California (16.2%), and Nevada (16.2%) had the highest age-adjusted prevalence of a history of foot ulcer among persons with diabetes, and Colorado (7.4%), Wisconsin (8.8%), and Hawaii (8.9%) had the lowest " (2).

Treatments for curing DFU are very far from optimal. Current standard of care can cure only about 30% of DFU and even the most advanced therapies, cell-based devices containing skin derived keratinocytes and fibroblasts, boosts the cure rate only to about 50%, leaving a tremendous unmet need for new effective cures for DFU, particularly in California. We anticipate that the development candidate that we propose, a stem cell-based “biological bandage”, will bring a new and effective cure to our citizens who are suffering from diabetic foot ulcers.

Sources: 1) California Health Care Survey, UCLA, http://www.chis.ucla.edu/
2) CDC reports Morbidity and Mortality Weekly Report (MMWR), http://www.cdc.gov/mmwr/preview/mmwrhtml/mm5245a3.htm

Funding Type: 
Disease Team Research I
Grant Number: 
DR1-01423
Investigator: 
Institution: 
Type: 
PI
Type: 
Co-PI
ICOC Funds Committed: 
$22 999 937
Disease Focus: 
Diabetes
Collaborative Funder: 
JDRF
Stem Cell Use: 
Embryonic Stem Cell
Cell Line Generation: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 

Diabetes exacts a tremendous toll on patients, their families, and society in general. Autoimmune Type 1 diabetes, often called juvenile-onset diabetes, is caused by a person’s own immune system mistakenly destroying their insulin-producing cells in the pancreas, known as beta cells. When those beta cells are lost, the ability to produce insulin in response to food intake is lost, and blood sugar can increase to toxic levels. Although not due to autoimmunity, Type 2 diabetics often lose their ability to produce insulin as well. While pharmaceutical insulin is commonly used to control both types of diabetes, it does not sufficiently replace beta cells, and the adverse short- and long-term effects of diabetes remain, including dangerous episodes of low blood sugar, nerve damage, blindness, kidney damage, foot ulcers leading to amputations, and cardiovascular disease. Ideally, one would like to replace lost beta cells, and attempts to do so have included the use of pancreatic transplants, beta cell (islet) transplants, and transplants of animal cells or tissues. Unfortunately, those approaches are hindered by 1) the limited amount of donor tissue available, and 2) issues regarding immunological complications between donors and recipients. To solve the first problem, the Diabetes Disease Team applying for this CIRM award has developed methods to make replacement beta cells from human embryonic stem cells (hESC), which can be reliably grown in large-scale batches. The hESC-derived beta cells have been shown to cure experimental diabetes in mice and rats. Regarding the issue of donor-recipient compatibility, the Team has had initial success with several strategies, including administering the cells inside a simple device, implantable under the skin, as well as next-generation pharmaceuticals that enable transplantation between unmatched individuals without major side effects.

With the critical proof-of-concept milestones behind us, the Team now needs to perform all of the manufacturing and laboratory testing required to assure reliable production of a safe and effective product, thereby generating the data needed to seek FDA approval to test the product in humans. The project engages over 30 scientists and physicians, as well as numerous associates and technicians, whose expertise covers all of the critical areas from process development and manufacturing to clinical testing of novel biomedical products. The proposal includes active project management, and regulatory and ethical oversight. The Team has well defined time lines and milestones to advance the candidate product to an FDA submission. If successful, testing in diabetic patients could begin as early as 3 years from the project initiation.

Statement of Benefit to California: 

Diabetes mellitus currently afflicts more than 250 million people worldwide, with projections of 380 million by the year 2030 (source: International Diabetes Federation). In 2007, there were an estimated 2.7 million Californians with diabetes (source: California Diabetes Program, California Department of Public Health). Further, the disease disproportionately affects certain minority groups and the elderly. Despite the use of insulin and advances in its delivery, the human cost of diabetes is underscored by the financial costs to society: tens of billions of dollars each year in California alone. The primary cause of Type 1 diabetes, and contributing significantly to Type 2 diabetes as well, is the loss of insulin-producing pancreatic beta cells. The proposed Disease Team will develop a beta cell replacement therapy for diabetes. If successful, the therapy will go beyond insulin function, and will perform the full array of normal beta cell functions, including responding in a more physiological manner than manual or mechanized insulin administration. Because they will be more physiological, the replacement cells should also reduce the long-term effects of diabetes. Moreover, the cell therapy will alleviate patients of the constant monitoring of blood glucose and painful insulin injections. For these reasons, it is possible that the product could transform the diabetes treatment landscape and replace pharmaceutical insulin in the market. This product will be available in California first, through clinical trials, and if approved by the FDA for commercial production, could eventually help hundreds of thousands of diabetic Californians. The product will substantially increase quality of life for diabetics and significantly reduce the health care burden in the state. The Team will employ various Californian physicians and scientists, and success of the Team will generate positive recognition for the state. Lastly, once commercially marketed, the product will yield additional jobs in manufacturing, sales, and related industries, and generate revenue for California. Given the market need and the clear feasibility, the product could become the most significant stem cell-based medical treatment of the coming decade, and that would be a great achievement for California, its taxpayers, and CIRM.

Progress Report: 
  • The CIRM Diabetes Disease Team is developing a cell therapy to treat insulin-dependent diabetes. The ultimate goal under CIRM Award DR1-01423 is to file an IND with the FDA to allow first-in-human clinical testing of the cell therapy product. To reach that goal, numerous research and development activities need to be successfully executed in parallel, and the project requires careful planning and agile management. This is particularly critical because the planned product is complex and, as a cutting-edge technology, extends into new regulatory territory. In Year 1 of this Award, virtually all aspects of the project remained on track and the 4-year time line to filing an IND remains the same.
  • The planned product is a combination therapy that is expected to alleviate a diabetic’s need to perform frequent blood monitoring and insulin injections. It will essentially replace or provide needed support to the endocrine pancreas that is lost or damaged in diabetes. The product consists of a human pancreatic progenitor cell population administered in a durable delivery device. Following administration, the progenitor cells mature into human pancreatic islets including functional insulin-producing glucose-responsive beta cells. Prototypes of the product have been tested in hundreds of rodents, and in proof-of-concept studies, this cell-device combination has cured rodents of drug-induced diabetes.
  • The pancreatic progenitor cells are manufactured from human embryonic stem (ES) cells through a series of precise steps in cell culture. Using ES cells as starting material allows for the mass production of progenitor cells that will be required if the product is successful, as ES cells are remarkably proliferative while still remaining stable. In Year 1 of the Disease Team Award, frozen cell banks of ES cells were manufactured under Current Good Manufacturing Practice (cGMP), as required to clinically test and commercialize a cell therapy. Additionally in Year 1, the specific details of the pancreatic progenitor cell manufacturing process were finalized, and documentation was put in place to allow cGMP manufacture of pancreatic progenitor cells for future animal and human studies.
  • The cell delivery device is a small flat sealed chamber made from a semi-permeable membrane. The device serves multiple purposes. It is intended to protect the cells from the patient’s immune system, which is particularly important in autoimmune (Type 1) diabetes. It retains the cells at the site of administration for ease of monitoring and possible removal if necessary. Importantly, while cells cannot pass through it, the semi-permeable membrane allows sugars, oxygen, and other nutrients in, to sustain and regulate the islet cells, and allows insulin and other endocrine proteins out, to regulate blood sugar and other metabolic physiology. In Year 1, numerous prototype configurations of the delivery device were tested in animals, and a final configuration was determined. A device manufacturing facility was designed and built. Manufacturing and testing equipment was installed, and documentation put in place for production of clinically compliant devices. As with cell manufacturing described above, device manufacturing can now proceed at a scale and level of quality that will facilitate pre-clinical and clinical testing of the combination product.
  • It is possible that the device alone will not be sufficient to protect the cells from a diabetic patient’s immune system. In anticipation of this possibility, the Diabetes Disease Team includes world-renowned immunologists who are establishing animal models to test and address this question accordingly. Fortunately, there are many pharmaceutical options, including cutting-edge technologies, that have proven effective in protecting transplanted human islets from immune rejection, and those might be applied to administration of this cell therapy product as well. The Disease Team’s clinical group is developing plans for the first-in-human testing and will incorporate a regimen of immune modulation as appropriate. In Year 1, the animal models to test the requirement for immune modulation and various regimens were established. In Year 2, these models will be used to investigate these questions.
  • The CIRM Diabetes Disease Team is developing a cell therapy to treat insulin-dependent diabetes. The ultimate goal under CIRM Award DR1-01423 is to file an IND with the FDA to allow first-in-human clinical testing of the cell therapy product. To reach that goal, numerous research and development activities need to be successfully executed in parallel, and the project requires careful planning and agile management. This is particularly critical because the planned product is complex and, as a cutting-edge technology, extends into new regulatory territory. In Year 2 of this Award, virtually all aspects of the project remained on track and the 4-year time line to filing an IND remains the same.
  • The planned product is a combination therapy that is expected to alleviate diabetes patients’ need to perform frequent blood monitoring and insulin injections. It will essentially replace or provide needed support to the endocrine pancreas that is lost or damaged in diabetes. The product consists of a human pancreatic progenitor cell population administered in a durable delivery device. Following administration, the progenitor cells mature into human pancreatic islets including functional insulin-producing glucose-responsive beta cells. Prototypes of the product have been tested in hundreds of rodents, and in proof-of-concept studies this cell-device combination has cured rodents of drug-induced diabetes.
  • The pancreatic progenitor cells are manufactured from human embryonic stem (ES) cells through a series of precise steps in cell culture. Using ES cells as starting material allows for the mass production of progenitor cells that will be required if the product is successful, as ES cells are remarkably proliferative while still remaining stable. In Year 1 of the Disease Team Award, frozen cell banks of ES cells were manufactured under Current Good Manufacturing Practice (cGMP), as required to clinically test and commercialize a cell therapy. In Year 2, these cGMP ES cell banks were tested to confirm that they performed similarly to previous banks. The cell manufacturing protocol was finalized and several batches of progenitor cells were manufactured to demonstrate the reliability of the protocol, in particular, with the new cGMP ES cells.
  • The cell delivery device is a small flat sealed chamber made from a semi-permeable membrane. The device serves multiple purposes. It is intended to protect the cells from the patient’s immune system, which is particularly important in autoimmune (Type 1) diabetes. It retains the cells at the site of administration for ease of monitoring and possible removal if necessary. Importantly, while cells cannot pass through it, the semi-permeable membrane allows sugars, oxygen, and other nutrients in, to sustain and regulate the islet cells, and allows insulin and other endocrine proteins out, to regulate blood sugar and other metabolic physiology. In Year 1, numerous prototype configurations of the delivery device were tested in animals, and a final configuration was determined. A device manufacturing facility was designed and built. Manufacturing and testing equipment was installed, and documentation put in place for production of clinically compliant devices. In Year 2, several batches of delivery devices were manufactured and tested under development phase-appropriate Quality Systems Regulations. A Good Laboratory Practice (GLP) study of the combination product, comprised of cells and devices manufactured with the newly developed systems, was performed to establish safety and efficacy in mice, prior to human testing. The results of the GLP study were favorable, suggesting the combination product will likely be safe and effective in the clinic.
  • It is possible that the device alone will not be sufficient to protect the cells from a patient’s immune system. In anticipation of this possibility, the Diabetes Disease Team includes world-renowned immunologists who are establishing animal models to test and address this question accordingly. In Year 2, preliminary data were collected using these animal models. The preliminary data suggest that the device will protect cells from autoimmunity.
  • In Year 3, the clinical protocol will be drafted, further refinements to product manufacturing including device loading will be established, and further pre-clinical testing will be performed. The Team plans to present the candidate product and development plans to regulatory agencies in order to obtain valuable feedback. The goal is to establish sufficient pre-clinical assurance to facilitate clinical testing at the end of the 4-year award period.
  • The CIRM Diabetes Disease Team is developing a stem cell-derived cell therapy to treat insulin-dependent diabetes. The ultimate goal under the 4-year CIRM Award DR1-01423 is to file an IND (Investigational New Drug application) with the FDA to allow first-in-human clinical testing of the cell therapy product. To reach this goal, numerous research and development activities need to be successfully executed in parallel. The project requires careful planning and agile management particularly because the planned product is complex and, as a cutting-edge technology, extends into new territory from a regulatory perspective. In Year 3 of this Award, all aspects of the project remained close to the original schedule. One study report from an external Contract Research Organization (CRO) was delivered two months later than planned, which delayed a meeting with the FDA and subsequent downstream activities. Accordingly, two months has been added to the original 4-year time line to filing an IND. The new target for IND filing is March 2014.
  • The planned product is a combination therapy that is expected to alleviate diabetes patients’ need to perform frequent blood monitoring and insulin injections. It will essentially replace or provide needed support to the endocrine pancreas that is lost or damaged in diabetes. The product consists of a human cell population containing pancreatic progenitors administered subcutaneously in a durable delivery device. Following administration, the progenitor cells mature into human pancreatic islet-like tissue including functional insulin-producing, glucose-responsive beta cells while in the device. Prototypes of the product have been tested in hundreds of rodents, and in proof-of-concept studies this cell-device combination has cured rodents of chemically-induced diabetes.
  • The pancreatic cells are manufactured from human embryonic stem (ES) cells through a series of precise steps in cell culture. In Year 3, a Cell Manufacturing Cleanroom was built and commissioned in preparation for manufacturing cells for clinical testing. Two new cell manufacturing formats, both amenable to the scale-up that will be required for commercial manufacturing, were also developed. At the end of Year 3, a Pilot Plant was established for process development and technology transfer of the cell manufacturing protocol.
  • The cell delivery device is essentially a small sealed envelope made from a semi-permeable membrane. It is expected to protect the cells from the patient’s immune system and retain the cells at the site of administration. At the same time it will allow sugars, oxygen, and other nutrients in, to sustain and regulate the islet-like tissue, and allow insulin and other endocrine proteins out, to regulate blood sugar and other metabolic physiology. In Years 1-2, prototype (‘animal-sized’) devices were produced and tested, the configuration was finalized, and a Device Manufacturing Facility with equipment for quality control testing was built. In Year 3, the clinical (‘human-sized’) device was designed and built. Also in Year 3, all ISO10993 (safety standards for medical devices from the International Organization for Standardization) testing was completed, establishing that the device and its component materials will be safe for human use. A prototype system to load the progenitor cells into the device was designed and built in Year 3. The Team established and staffed a Combination Product Group.
  • In Year 3, a GLP (good laboratory practice) study of the combination product to test safety and efficacy in mice, prior to human testing, was completed by an independent CRO. The results were favorable, providing further rationale for advancement of the product into clinical testing.
  • To evaluate the potential of the device to protect the implanted cells from a patient’s immune system, the Diabetes Disease Team includes world-renowned immunologists who are establishing animal models to test and address this question. In Year 3, animal studies demonstrated that the device indeed protects animal cells from allo-immunity (addressing differences between donor and recipient tissues), suggesting the human pancreatic cells will also be protected in the product planned for human use.
  • In Year 3, the Team met with the FDA in a Pre-IND meeting. This meeting was informative and provided clarity on the remaining activities before an IND can be submitted and a clinical trial initiated.
  • During Years 1-3, the clinical protocol was drafted, and in Year 4 it will be finalized while the clinical sites are prepared. Also in Year 4, refinements to product manufacturing including device loading will be established, and additional pre-clinical testing will be performed to further assure safety of all aspects of the clinical plan. The goal is to establish a body of pre-clinical data that supports clinical testing at the end of the 4-year award period.
  • In the years just prior to the establishment of the CIRM Diabetes Disease Team, scientists at ViaCyte (then known as Novocell) clearly demonstrated in mice the great promise of using pancreatic progenitor cells as a potential cell replacement therapy for type 1 diabetes.
  • In type 1 diabetes, the insulin-producing cells of the pancreas are lost to autoimmunity. ViaCyte has shown that human pancreatic progenitors, derived from human embryonic stem cells (hESC) in culture, will further mature to human pancreatic islet cells including glucose-responsive insulin-producing cells after implantation, and that these cells can rescue or protect mice from experimentally induced diabetes [Kroon et al., 2008, Nature Biotechnology, 26(4): 443-452]. Further, the group demonstrated that the pancreatic progenitor cells could effectively protect mice from experimental diabetes when implanted in a “macro-encapsulation” device, which is designed to protect the cells from allo- and auto-immunity.
  • In short, the ViaCyte team developed a procedure and a strategy to replace the insulin-producing cells lost in type 1 diabetes, leveraging the great potential of hESC as an approach to large-scale production of replacement cells, and macro-encapsulation as a way to avoid immunosuppressant drugs typically needed in same-species cell and organ transplantation. The next crucial steps were to translate this promising research into a clinically acceptable process, assure the safety and efficacy of the approach in independent animal studies, establish a plan to test in patients with type 1 diabetes, and submit the complete data package to the regulatory authorities, including the FDA, in order to allow clinical trials to commence.
  • Over the past four and half years, the CIRM Diabetes Disease Team achieved the goals and milestones that it proposed at its inception in 2009. Substantial progress was made in advancing the stem cell-derived cell therapy and delivery device combination product from research phase to clinical development, including initiation of a Phase 1/2 clinical trial upon completion of this work.
  • In cell product development, the achievements included manufacture of Master and Working Cell Banks of the specific hESC starting material under Good Manufacturing Practices (GMP), development of a robust, reliable, scalable manufacturing process for differentiation of hESC into pancreatic progenitor cells (PEC-01™ cells), and development of cryopreservation and thawing and recovery methods for preparation of PEC-01 cells prior to loading into macro-encapsulation (Encaptra®) devices.
  • In device development, achievements included assessing and establishing materials and methods, and formalizing procedures for manufacturing Encaptra devices. Devices and their materials were thoroughly tested for biocompatibility and safety under ISO 10993 regulations. Custom manufacturing and testing methods and protocols were established.
  • In parallel with cell and device development, the team established custom materials and methods for combining these two main components into the product candidate (VC-01™ product). This included aseptic processes for loading cells into devices, sealing the devices, and placing them into custom packaging for delivery to the clinic.
  • Extensive Quality Control and Quality Assurance (QC/QA) systems were designed and implemented to assure standardized, reliable, safe and efficacious VC-01 product would be produced for clinical research. As biologicals (the cells) and devices fall under different regulations, the team needed to develop a custom hybrid quality management system that addressed both sets of regulations.
  • Numerous pre-clinical studies were performed in preparation for clinical testing, including three safety and efficacy studies of the VC-01 combination product under Good Laboratory Practices (GLP) at independent contract research organizations. Scientists in the immunology laboratories at University of California, San Francisco, and the La Jolla Institute for Allergy and Immunology further examined and demonstrated the utility of macro-encapsulation to protect implanted cells from allo- and auto-immunity in several animal models. The pre-clinical studies collectively indicated that proceeding to human testing was warranted and appropriate.
  • In the final years of the project, a Phase 1/2 first-in-human clinical trial was prepared. The trial is designed to provide critical insights into safety and the potential efficacy of the product concept. Lastly, the team had successful interactions with regulatory authorities, including a pre-IND submission and meeting with the FDA, and as a culmination of all of the work, a device master file (MAF) and investigational new drug application (IND) were submitted to the FDA. In the last month of the project the regulatory documents were accepted by the FDA, and the path was set to commence clinical testing of the product in patients with type 1 diabetes.
Funding Type: 
Basic Biology IV
Grant Number: 
RB4-06144
Investigator: 
Type: 
PI
ICOC Funds Committed: 
$1 391 400
Disease Focus: 
Diabetes
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 

The loss of pancreatic beta-cells in type 1 diabetes results in absence of insulin secreted by the pancreas, and consequently elevated blood sugar which leads to various long-term complications. Diabetic patients would benefit tremendously from availability of transplantable replacement beta-cells. Much of current research focuses on producing beta-cells from stem cells. Despite some progress, it is at present still not possible to generate functional beta-cells in culture. The beta-like cells generated with current protocols in vitro lack key features of normal beta-cells, most notably the ability to secrete insulin a regulated manner. However, when stem cell-derived beta-cell precursors are transplanted into mice, they acquire properties of functional beta-cells, indicating that the precursors have the potential to transition into a mature beta-cell state.

By comprehensively comparing the molecular profiles of mature, functional beta-cells and malfunctioning insulin-producing cells generated in vitro, we have identified molecular cues that are not appropriately induced under current culture conditions. These studies have led to short list of candidate regulators of beta-cell maturation. We propose to manipulate these candidate factors in stem cell-derived beta-cell precursors in culture, with the goal of forcing them to adopt a mature phenotype. We will first characterize these cells in vitro and then test functionality in diabetic animal models.

Statement of Benefit to California: 

Diabetes is a metabolic disorder that affects 8.3% of the U.S. population. Average medical expenditures among people with diabetes are 2.3 times higher than those of people without diabetes. The disease is characterized by either absolute insulin deficiency due to the autoimmune destruction of pancreatic insulin-producing beta-cells [Type 1 diabetes], or relative insulin deficiency due to defective insulin secretion or insulin sensitivity [Type 2 diabetes]. The resulting elevated blood glucose levels eventually lead to damage of the blood vessels followed by kidney failure, blindness, neuropathy, heart disease, and stroke. Despite current treatment regimens of several insulin injections per day, blood glucose levels still fluctuate significantly in diabetic patients, making diabetes the seventh leading cause of death in the United States. Alternative approaches to insulin injections include attempts to develop a cell therapy by producing transplantable beta-cells from stem cells. A cell therapy would lead to better blood glucose control and therefore ameliorate long-term complications. This proposal seeks to identify factors that force stem cell-derived beta-cells to functionally mature in culture with the goal to produce an unlimited source of transplantable beta-cells. Given the high prevalence of diabetes in California, we believe that the proposed research will have tremendous benefit to the State of California and its citizens.

Progress Report: 
  • The loss of pancreatic beta-cells in type 1 diabetes results in absence of insulin secreted by the pancreas, and consequently elevated blood sugar which leads to various long-term complications. Diabetic patients would benefit tremendously from availability of transplantable replacement beta-cells. Much of current research focuses on producing beta-cells from stem cells. Despite some progress, it is at present still not possible to generate functional beta-cells in culture. The beta-like cells generated with current protocols in vitro lack key features of normal beta-cells, most notably the ability to secrete insulin a regulated manner. However, when stem cell-derived beta-cell precursors are transplanted into mice, they acquire properties of functional beta-cells, indicating that the precursors have the potential to transition into a mature beta-cell state.
  • This proposal explores strategies for maturing beta-cell precursors in the culture dish with the goal to produce fully functional insulin-producing beta-cells in vitro. Previous studies from our laboratory have resulted in a short list of candidate regulators of beta-cell maturation. We propose to manipulate these candidate regulators in vitro in order to force beta-cell precursors to adopt a mature phenotype. We have now established a robust in vitro system for culturing and manipulating beta-cell precursors. We have also generated and tested requisite reagents for manipulating precursors in the culture dish. Over the next year, we will obtain first results from these manipulations.
  • The potential to generate functional pancreatic beta cells from human embryonic stem cells provides a promising avenue for beta cell replacement therapy for treatment of diabetes mellitus. Despite the rapid advancements in the field, current protocols still do not produce fully functional beta cells and the production of these cells takes at least six weeks. Understanding the molecular cues that regulate how a beta cell develops and matures would be critical in improving current approaches to generate functional beta cells. In our lab, we found that hundreds of genes enriched in functional beta cells were not properly induced in the nonfunctional cells derived from embryonic stem cells, suggesting that manipulating critical regulators that can affect multiple beta cell genes simultaneously might be instrumental in directing beta cell differentiation and maturation in the culture dish. Our studies have identified several novel regulators of gene expression, including transcription factors and small non-coding RNAs, which we predict will have critical roles in beta cell maturation. These novel regulators are highly expressed in insulin-producing islets and have very low expression in non-functional beta-like cells. This suggests that forcing expression of these regulators could accelerate the formation of functional mature beta cells. Preliminary studies so far show that forced expression of select candidates in immature precursor cells can induce the expression of several genes critical for beta cell maturation. Over the next year, we will determine, by manipulating the expression of our candidate regulators, the optimal conditions required for inducing the production of mature beta cells in vitro.
Funding Type: 
Basic Biology III
Grant Number: 
RB3-02266
Investigator: 
Type: 
PI
ICOC Funds Committed: 
$1 313 649
Disease Focus: 
Diabetes
Stem Cell Use: 
Embryonic Stem Cell
oldStatus: 
Active
Public Abstract: 

There remains an urgent and critical need for a cell-based cure of diabetes, one of the most costly diseases in California. Islet transplantation with persistent immune suppression has shown promise in curing type 1 diabetes (TID). However, one major obstacle towards large scale implementation of this approach is the shortage of engraftable islets. Human ES cells (hESCs), which can undergo unlimited self-renewal and differentiate into all cell types in the body, have the potential to become an unlimited source of pancreatic β cells. Significant challenges, including the lack of chemical defined conditions for reproducibly differentiating hESCs into endocrine precursors (EPs), lack of strategy to purify these EPs to avoid teratoma risk, and destruction of engrafted islets by allogeneic and autoimmune rejection despite persistent immune suppression, hinder clinic development of this promising hESC based therapy.

Ongoing research in our laboratories is directed at developing novel strategies to derive β-cells from hESCs. Of the several genetic factors that contribute to stem cells differentiation, miRs (microRNAs) are emerging as important determinants. We hypothesize that identification and validation of the temporal expression of miRs at discrete, functionally defined and genetically marked stages of hESC differentiation to insulin-producing cells, when combined with a computational/systems biology approach, will create a population of cells of significant therapeutic impact. The proposed studies will translate basic large-scale analysis of miR and mRNA from pancreatic precursors derived from hESC into a fundamental understanding of differentiation. This in turn will ultimately lead to novel treatments for T1D.

In this project we will elucidate the importance of miRs in pancreatic cell differentiation through functional testing, genetic marking, deep sequencing, computational analysis, and validation. Within the context of the above-stated general aims the sequencing studies will be initiated for 3 reasons: 1) to establish on site the most powerful approaches currently available for measuring gene identity and expression 2) to ensure that novel and established miRs are evaluated for changes in expression during hESC differentiation 3) to validate targets of miR action. Application of this emerging technology to β-cell genesis will allow the generation of miR and mRNA profiles from uniform cell populations and validation through functional assays. Together, this information will help to better understand, describe, and ultimately optimize hESC differentiation. Basic research from this project has the potential to create a paradigm shift in understanding the cellular ontogeny of the pancreas and help identify which cell types can be used for transplantation therapy in T1D.

Statement of Benefit to California: 

Diabetes has devastating consequences on both those afflicted and on State/National healthcare costs, and, given the staggering rise in both occurrence and costs, diabetes alone possesses the potential to completely overwhelm our healthcare system. There remains an urgent and critical need for a cell-based cure. In 2007, diabetes directly affected 1 in 10 Californians (2.7 million), costing the state $24.5B annually. There have been documented, significant increases in the occurrence of both type 1 and type 2 diabetes in youths under 18 years of age (0.16% of youth <18 yr have type 1 diabetes nationally). There are more than 7,000 diabetic children within [REDACTED] alone.

The following alarming statistics are provided by the California Department of Public Health, California Diabetes Control Program, CDC and NIH/NIDDK:
• In the U.S., diabetes is the most costly chronic disease, costing $132B annually. This is predicted to rise to $192B by 2020.
• Nearly 1 in 3 Medicare dollars and 1 in 10 of U.S. healthcare dollars are spent treating diabetes.
• Diabetics average $13,243/year in health care costs, 2.4 times more than non-diabetics.
• 7% of the US population has diabetes.
• Every 24 hours, 4,100 Americans are diagnosed with diabetes, 613 American diabetics die of the disease and another 55 go blind.
• Worldwide, every 10 seconds a diabetic dies and two new people develop diabetes.
• Worldwide expenditures on insulin alone are estimated to be $15 billion annually and growing.

This research would benefit the State of California and its citizens on multiple fronts. First and foremost, positive results will create a new development candidate for cell-based therapy for type 1 diabetes with the potential for avoiding the risk of tumor formation - a consequence that hinders the development of any human ES cell based therapy. Second, the application of new technologies would enhance the prospects for new biological agents that will require scale up efforts not available to academics. The creation of progenitor cells for any chronic disease, diabetes in our case, will enhance the prospects for the increase in personnel at the scientific and technical level for both academic labs and biotech companies. Finally, this work may obviate the need for immune suppression therapy that today carries serious side effects including propensity to infections and cancer, abnormalities in lipid metabolism and hypertension, and even damage to the transplanted cells as it occurs following islet transplantation procedures, the only available therapy nowadays for insulin-dependent diabetes. Avoidance of these complications represents a significant positive step in the reduction of health care expenses directly attributed to diabetes and its complications.

Progress Report: 
  • The long term goal of our research is to understand the biochemical processes that regulate differentiation of human embryonic stem cells (hESCs) into pancreatic progenitor cells, and ultimately, glucose-responsive, insulin producing (beta) β cells. Islet transplantation with persistent immune suppression has shown promise in curing type 1 diabetes (TID). However, one major obstacle towards large scale implementation of this approach is the shortage of engraftable islets. hESCs, which can undergo unlimited self-renewal and differentiate into all cell types in the body, have the potential to become an unlimited source of pancreatic β cells, however, significant challenges have hindered clinic development of this promising hESC based therapy.
  • Ongoing research in our laboratory is directed at deriving β-cells from hESCs. Of the several genetic factors that contribute to stem cells differentiation, miRNAs (microRNAs) are emerging as important determinants. miRNAs are noncoding, regulatory RNAs expressed dynamically during differentiation of hESC. Mapping developmental expression of miRNAs during transition from pluripotency to pancreatic progenitors will help clarify the mechanisms underlying lineage specification and ultimately enhance differentiation protocols. Specifically, the objectives of this CIRM grant are to elucidate the role miRNAs play in the development of hESC into cells of endocrine lineage and to provide crucial details on the molecular architecture of endocrine precursor populations, lineage specification, and β-cell maturation.
  • The central hypothesis driving the research is that miRNAs are essential regulators of endocrine cell development. We are working under the postulate that miRNAs are logical targets for in vitro experimentation because of their role in mediating pancreatic cell development. Our aims are as follow:
  • Aim 1 - Generate miR expression profiles using deep sequencing for defined stages of development from pluripotent to endocrine cells and select candidate miRs for manipulations involving silencing and overexpression.
  • Aim 2 - Identify miRs targets through deep sequencing of RNA induced silencing complexes (RISC) in defined cell populations and assessment of their roles in differentiation in vitro and after experimental transplantation.
  • During the current funding period, progress has been made on both specific aims originally proposed. From this work, one manuscript and one review article have been published and two other research articles are submitted/in review.
  • Published studies. A) “The SDF-1α/CXCR4 axis is required for proliferation and maturation of human fetal pancreatic endocrine progenitor cells.” was published in PLoSONE. B) “From pluripotency to islets: miRNAs as critical regulators of human cellular differentiation” was published in Advances in Genetics.
  • Submitted studies. A) “sRNA-seq analysis of human embryonic stem cells and definitive endoderm reveal differentially expressed microRNAs and novel isomiRs with distinct targets” is in revision at Stem Cells. B) “Jak/Stat and MAP kinase signaling regulate human embryonic stem cell pluripotency” will be re-submitted to Cell Stem Cell in early October.
  • Work in progress. A) Deep sequencing of miRNAs from a purified population of PDX-1+ cells derived from hESC. Towards our goal of understanding the role miRNAs play in driving differentiation of insulin producing cells from pluripotent hESC, we have sequenced miRNAs from a heterogeneous population of hESC that have been directed towards endocrine lineage. B) Deep sequencing of miRNAs at 24 hour intervals during hESC differentiation towards pancreatic precursors. A major undertaking during the first year of funding is to sequence the changes in miRNA expression at selected intervals during the differentiation process. This information is critical for us to develop algorithms to determine how miRNAs drive differentiation and for identification of miRNA/mRNA targets. C) Development of algorithms to analyze change in miR expression in complex systems. During the first phases of the CIRM project, Natural Selection Inc. focused on algorithms to analyze change in microRNA expression over multiple data sets. D) Generation of a population of PDX1+ cells using zinc finger nuclease technology. One critical goal of the proposed studies is to generate a purified population of endocrine precursor cells. Although some technical problems with construction of the vector arose, we believe that we have overcome the major obstacles and will have these cells for microRNA analysis during the next funding period.
  • Together, the information generated in this study is helping us to better understand, describe, and ultimately optimize hESC differentiation. We believe that the results from this project have the potential to create a paradigm shift in understanding the cellular ontogeny of the pancreas and help identify which cell types can be used for transplantation therapy in T1D.
  • The long term goal of our research is to understand the biochemical processes that regulate differentiation of human embryonic stem cells (hESCs) into pancreatic progenitor cells, and ultimately, glucose-responsive, insulin producing (beta) β cells. hESCs, which can undergo unlimited self-renewal and differentiate into all cell types in the body, have the potential to become an unlimited source of pancreatic β cells, however, significant challenges have hindered clinic development of this promising hESC based therapy.
  • Ongoing research in our laboratory is directed at deriving β-cells from hESCs. Of the several genetic factors that contribute to stem cells differentiation, miRNAs (microRNAs) are emerging as important determinants. miRNAs are noncoding, regulatory RNAs expressed dynamically during differentiation of hESC. Mapping developmental expression of miRNAs during transition from pluripotency to pancreatic progenitors will help clarify the mechanisms underlying lineage specification and ultimately enhance differentiation protocols. Specifically, the objectives of this CIRM grant are to elucidate the role miRNAs play in the development of hESC into cells of endocrine lineage and to provide crucial details on the molecular architecture of endocrine precursor populations, lineage specification, and β-cell maturation.
  • The central hypothesis driving the research is that miRNAs are essential regulators of endocrine cell development. We are working under the postulate that miRNAs are logical targets for in vitro experimentation because of their role in mediating pancreatic cell development. Our aims are as follow:
  • Aim 1 - Generate miR expression profiles using deep sequencing for defined stages of development from pluripotent to endocrine cells and select candidate miRs for manipulations involving silencing and overexpression.
  • Aim 2 - Identify miRs targets through deep sequencing of RNA induced silencing complexes (RISC) in defined cell populations and assessment of their roles in differentiation in vitro and after experimental transplantation.
  • During the current funding period, progress has been made on both specific aims originally proposed.
  • Published studies. “A) “Imaging human fetal pancreas.” was published in Journal of Visualized Experiments.
  • Provisional Patents Filed. A) “Novel combinations of transcriptional gene regulators”. B) “Assay to detect onco-miRs circulating in serum or cells”
  • Work in progress.
  • A) Paired microRNA expression and development of a reporter system for lineage fate.
  • We have made a cell line that reports the expression of PDX1, a marker for both pancreatic precursors and for mature beta cells, in order to select and purify the target cells in late differentiation. miR-375 was previously described by our lab to be the most abundant miRNA in definitive endoderm (DE). Other labs have shown that miR-375 is also expressed in pancreatic during development, and specifically in beta cells in mature islets, where it regulates insulin secretion. Conversely, miR-122 is the most highly expressed miRNA in liver, which arises in the region of endoderm that is closest to the pancreatic buds. In our hESC differentiation protocol, only miR-375 is expressed at the DE stage, which is typically about 98% pure. miR-122 is not expressed in DE, but increases in levels to coincide with lower miR-375 expression as a more heterogeneous mixture of cells form as DE differentiates into multiple lineages. We have generated a reporter cell lines that can distinguish pancreatic cells from liver cells in post-DE differentiation, and possibly mature beta cells from other endocrine cells.
  • B) Deep sequence purified hESC populations from selected time points during hESC differentiation and develop algorithms to analyze change in miRNA expression in complex systems.
  • In our previous approach, we applied pattern filters to the data to see which miRs matched a particular filter. This was valuable as it helped us determine which miRs had similar expression patterns. However there was still a lot of variance. Therefore, we wre-did the analysis using a new clustering methods developed in conjunction with NSI. The latest approach has given us very valuable insight into the data. We have found that master regulators of miRs exist and/or are being regulated by another regulator but that regulator is biasing them over a large time scale (weeks). The day-to-day fluctuations that most investigators focus upon are not found in this latest group, suggesting that short-term and long-term miR regulation are differentially regulated.
  • Together, the information generated in this study is helping us to better understand, describe, and ultimately optimize hESC differentiation. We believe that the results from this project have the potential to create a paradigm shift in understanding the cellular ontogeny of the pancreas and help identify which cell types can be used for transplantation therapy in T1D.
  • The long term goal of our research is to understand the biochemical processes that regulate differentiation of human embryonic stem cells (hESCs) into pancreatic progenitor cells, and ultimately, glucose-responsive, insulin producing (beta) β cells. Islet transplantation with persistent immune suppression has shown promise in curing type 1 diabetes (TID). However, one major obstacle towards large scale implementation of this approach is the shortage of engraftable islets. hESCs, which can undergo unlimited self-renewal and differentiate into all cell types in the body, have the potential to become an unlimited source of pancreatic β cells, however, significant challenges have hindered clinic development of this promising hESC based therapy.
  • Ongoing research in our laboratory is directed at deriving β-cells from hESCs. Of the several genetic factors that contribute to stem cells differentiation, miRNAs (microRNAs) are emerging as important determinants. miRNAs are noncoding, regulatory RNAs expressed dynamically during differentiation of hESC. Mapping developmental expression of miRNAs during transition from pluripotency to pancreatic progenitors will help clarify the mechanisms underlying lineage specification and ultimately enhance differentiation protocols. Specifically, the objectives of this CIRM grant are to elucidate the role miRNAs play in the development of hESC into cells of endocrine lineage and to provide crucial details on the molecular architecture of endocrine precursor populations, lineage specification, and β-cell maturation.
  • The central hypothesis driving the research is that miRNAs are essential regulators of endocrine cell development. We are working under the postulate that miRNAs are logical targets for in vitro experimentation because of their role in mediating pancreatic cell development. Our aims are as follow:
  • Aim 1 - Generate miR expression profiles using deep sequencing for defined stages of development from pluripotent to endocrine cells and select candidate miRs for manipulations involving silencing and overexpression.
  • Aim 2 - Identify miRs targets through deep sequencing of RNA induced silencing complexes (RISC) in defined cell populations and assessment of their roles in differentiation in vitro and after experimental transplantation.
  • Published studies. A) “sRNA-seq analysis of human embryonic stem cells and definitive endoderm reveal differentially expressed microRNAs and novel isomiRs with distinct targets”
  • Submitted studies. Recently, we submitted a manuscript entitled “MicroRNA Dynamics During Human Embryonic Stem Cell Differentiation to Pancreatic Endoderm” to Stem Cells and Development. This work is a comprehensive microRNA study that details our deep sequencing experiments. In the study, we describe changes in microRNA expression that occur during the first 10 days of pancreatic progenitor formation, confirm expression of selected miRNAs at various time points, correlate expression with known mRNA targets, and integrate protein expression with data from our quantitative LC/MS experiments. This work provides a rare, simultaneous and comprehensive look at hESC for the microRNA, mRNA, and protein level.
  • Work in progress.
  • A) Transcriptional Regulation by Ago2 and miRNAs.
  • microRNAs and hESC differentiation. MicroRNAs (miRs) regulate post-transcriptional gene networks and function in a manner analogous to transcription factors. Mature miRNAs are partially complementary to one or more messenger RNAs (mRNA), and function primarily to down regulate gene expression. The importance of miRNA activity in hESC during mammalian development has been established by deleting genes necessary for global miRNA biogenesis. Our recent work has found that miRNA expression in hESC is dynamic, indicating that miRNAs drive differentiation. We have been working under the hypothesis that transcription of miRNA loci are driven by mature miRNA and Ago2, and by manipulating Ago2, we can effect levels of precursor and mature miRNA expression and ultimately regulate hESC differentiation.
  • B) PKCβ isozyme expression is regulated by miR-653 during Definitive Endoderm Formation.
  • Recently, we have employed deep sequencing to generate temporal maps of changes in microRNA expression during the first ten days of hESC differentiation towards endocrine cell development. The results from the miRNA/mRNA expression studies identified the beta isozyme of PKC as a potential target of miR-653. We found that miR-653 is specifically associated with DE fate and contains a strong consensus binding site in the 3’ UTR of PKCβ. In both Cyt49 and H9 hESC lines, the log2 value for mRNA expression of PKCβ dramatically dropped during DE formation. This was unique to the beta PKC isoform. Western blot analysis of endogenous PKCβII expression confirmed loss at the protein level as cells left pluripotency for DE.
Funding Type: 
Early Translational I
Grant Number: 
TR1-01277
Investigator: 
Name: 
Type: 
PI
ICOC Funds Committed: 
$5 165 028
Disease Focus: 
Diabetes
Stem Cell Use: 
iPS Cell
Cell Line Generation: 
iPS Cell
oldStatus: 
Closed
Public Abstract: 

Human embryonic stem cells (hESCs) can undergo unlimited self-renewal and differentiate into all the cell types in the human body, and thus hold great promise for cell replacement therapy. However, one major problem for hESC-based therapy is that the cells derived from hESCs will be rejected by the recipient and can only be tolerated under persistent immunosuppression, which itself can cause cancer and infection. Recent development of induced pluripotent stem cells (iPSCs), which are generated from somatic cells of individual patient with defined factors and very similar to hESCs, could provide ideal cell source for transplantation by avoiding graft rejection in the patient. In addition, the disease-specific iPSCs can be used as human disease models for more reliable testing of the efficacy and toxicity of drugs. However, there are several major bottlenecks that prevent the development of iPSCs in human therapy and drug discovery. The overall goal of this proposal is to resolve the major bottlenecks remained in human iPSC biology to make it feasible for human therapy and drug discovery. We propose to develop safe and efficient approach to generate iPSCs from human patients. We propose to develop strategies to eliminate the risk of teratomas associated with the undifferentiated iPSCs. We propose to develop mouse model with functional human immune system to study the immune responses and tolerance during transplantation. Resolving these bottlenecks will greatly facilitate the development of hESCs into stem cell therapy and disease models for drug discovery.

Statement of Benefit to California: 

Diabetes and heart diseases remain the most costly diseases in our State and Nation. In the case of diabetes, 1 of every 10 Californians (2.7 million) were afflicted with diabetes in 2007, costing the State $24.5 billion annually. There is a significant increase in the occurrence of both types of diabetes in youths under 18 years of age (0.16% of youth <18 yr have type 1 diabetes nationally). Simply put, diabetes is having devastating consequences on both those afflicted and on State/National healthcare costs, and, given the staggering rise in both occurrence and costs, diabetes possesses the potential to completely overwhelm our healthcare system. There remains an urgent and critical need for a cell-based cure of diabetes. There is hope, since transplantation of functional β cells from human donors has been validated clinically to cure diabetes.

While significant progress has been made in the derivation of functional β cells and cardiomyocytes from human ES cells, these allogenic cells will be rejected by the recipient upon transplantation unless the immune system of the recipient is persistently suppressed. However, immune suppression itself has severe consequences with significantly increased risk of cancer and infection. This problem might be resolved by the recent breakthrough in induced pluripotent stem cell (iPSCs), which can be reprogrammed from somatic cells of human patients by defined factors and thus can provide a renewable source of autologous cells for transplantation. In addition, the disease-specific iPSCs will provide the much needed disease models to more reliably predict the drug responses in humans. With our significant progress in producing iPSCs without viral vectors or permanent genetic modification, our proposed research will resolve the major bottlenecks that hinder the development of iPSCs into human therapy and drug discovery. If successful, the funding spent now on research is nominal when compared to the billions that will be saved in treatment costs and the improved quality of life for patients.

Progress Report: 
  • Human induced pluripotent stem cells (hiPSCs), reprogrammed from somatic cells with defined factors, are similar to human ES cells (hESCs) and could provide ideal cell source for transplantation by avoiding immune rejection. In addition, disease-specific hiPSCs could provide improved disease models to predict drug responses in humans. The permanent genetic modification by random viral integration and spontaneous reactivation of reprogramming factors lead to cancer risk and abnormal differentiation. During the past year, we have made progresses to develop a combination of chemical and episomal approaches to reprogram human cells into iPSCs without genetic modifications. We have developed the constructs for the pre-transplant strategies to eliminate the teratomas risk of undifferentiated iPSCs. We have started to improve conditions for iPSC differentation into beta cells. In addition, we developed mouse models reconstituted with human immune system to enable us to study the immunogenicity and tolerance of cells derived from isogenic iPSCs.
  • During the past year, we have made significant progress in the proposed research. One most important finding is the discovery of the immunogenicity of the cells derived from induced pluripotent stem cells (iPSCs). This immunogenicity is due to the abnormal gene expression during the differentiation of iPSCs. This finding, published in the journal Nature, indicates that we need to perform more research on iPSCs before moving forward into clinical trial. Another major finding is the discovery of a safer way to improve the efficiency of iPSC production. In addition, we have made some progress in developing a genetic approach to eliminate the teratomas risk associated with the undifferentiated pluripotent stem cells.
  • During the past funding period, we have accomplished the established milestones. We have compared the genomic stability of iPSCs generated with various approaches. We have developed a genetic approach to eliminate the teratomas risk associated with undifferentiated pluripotent stem cells. We have evaluated the immunogenicity of cells derived from human iPSCs.
  • We have achieved the milestones and completed the proposed research during the no-cost extension period.

Pages