Designing monitoring systems for beta-cell differentiation from ES cells

Funding Type: 
SEED Grant
Grant Number: 
RB2-01645
ICOC Funds Committed: 
$0
Stem Cell Use: 
Embryonic Stem Cell
iPS Cell
Public Abstract: 
Type I diabetes (TID) patients suffer from insufficient or lack of insulin to regulate blood glucose levels. Due to loss of pancreatic beta-cells, the only treatment for TID patient is transplantation of donor islets. With the improved transplantation technique, it still requires 2-4 donor pancreas for each successful transplantation. The shortage of donor islets and inefficiency of beta-cell regeneration presented an urgent need for alternative sources of beta-cells. Embryonic stem (ES) cell-derived insulin producing cells are regarded as a possible means to overcome these limitations. However, difficulties in controlling the fate of ES cell differentiation have presented significant challenge for using ES cells to treat type I diabetes. Current protocols of ES cell differentiation to beta-cells rely on trying different combinations of medium factors. A lack of monitoring system has made the effort to improve the beta-cell proliferation protocol a blind search. In this grant proposal, we intend to design a monitoring system to monitor the differentiation process. This monitoring system is based on the physiological development of the pancreas. During pancreas development, several distinct stage with different cell markers have been identified to be critical. Using these markers as a monitoring guide will aid in our effort of improving beta-cell differentiation protocol. In this study, we propose to design fluorescent reporter markers to monitor the appearance and disappearance of these markers. The system designed here will be useful tools for future studies targeted at improving the current beta-cell differentiation protocol. A efficient and reliable beta-cell differentiation protocol is the critical step toward using ES cell based therapy for the treatment of type I diabtetes.
Statement of Benefit to California: 
The proposed study is an important step towards establishing a reliable, robust differentiation system for beta-cells. The percentage of population who has diabetes in California is among the top in the 50 states. Therefore, a treatment targeted at diabetes will be of significant benefit to the California citizens. For type I diabetes for whom transplantation is the only reliable treatment, an alternative treatment is urgently needed and is of significant benefit to the Californians.
Progress Report: 
  • The ultimate goal of the proposed study is to identify approaches to increase the production of therapeutically useful blood cells from human ESCs and patient-specific iPSCs. Currently, bone marrow transplantation is the best way to cure many blood-related disorders, such as sickle cell anemia, thalassemia, and blood cancers like leukemia. Furthermore, blood transfusion is an effective way to rapidly counteract blood cell loss due to ablative treatments, such as chemotherapy and radiation therapy. Unfortunately, the limiting factor in transplantation and transfusion treatments is the lack of matched donors. The ability to producing unlimited numbers of blood stem cells and/or functioning differentiated blood cells from human ESCs and patient-derived iPSCs will greatly improve the opportunity of such treatments. Transcription factors play important roles in regulating cell proliferation and differentiation. RUNX1 is a transcription factor that is expressed in blood cells and regulates the expression of many blood cell related genes. Therefore, the specific aim of our studies is to examine the effect of RUNX1 on blood cell formation, expansion, and differentiation from human embryonic stem cells and induced pluripotent stem cells. During the first year of funding period, we have established the cell culture and differentiation systems in our laboratory, generated necessary DNA constructs for the proposed studies, and produced transcription factors for testing the effect.
  • The ultimate goal of the proposed study is to identify approaches to increase the production of therapeutically useful blood cells from human ESCs and patient-specific iPSCs. Currently, bone marrow transplantation is the best way to cure many blood-related disorders, such as sickle cell anemia, thalassemia, and blood cancers like leukemia. Furthermore, blood transfusion is an effective way to rapidly counteract blood cell loss due to ablative treatments, such as chemotherapy and radiation therapy. Unfortunately, the limiting factor in transplantation and transfusion treatments is the lack of matched donors. The ability to producing unlimited numbers of blood stem cells and/or functioning differentiated blood cells from human ESCs and patient-derived iPSCs will greatly improve the opportunity of such treatments. Transcription factors play important roles in regulating cell proliferation and differentiation. RUNX1 is a transcription factor that is expressed in blood cells and regulates the expression of many blood cell related genes. Therefore, the specific aim of our studies is to examine the effect of RUNX1 on blood cell formation, expansion, and differentiation from human embryonic stem cells and induced pluripotent stem cells. During the second year of funding period, we have performed hematopoietic cell differentiation using both human ESCs and iPSCs in the presence and absence of this transcription factor. Our results indicate that this factor promotes the production of blood stem cells and progenitors. We will make additional amount of this factor and further confirm our initial finding.
  • The ultimate goal of the proposed study is to identify approaches to increase the production of therapeutically useful blood cells from human ESCs and patient-specific iPSCs. Currently, bone marrow transplantation is the best way to cure many blood-related disorders, such as sickle cell anemia, thalassemia, and blood cancers like leukemia. Furthermore, blood transfusion is an effective way to rapidly counteract blood cell loss due to ablative treatments, such as chemotherapy and radiation therapy. Unfortunately, the limiting factor in transplantation and transfusion treatments is the lack of matched donors. The ability to producing unlimited numbers of blood stem cells and/or functioning differentiated blood cells from human ESCs and patient-derived iPSCs will greatly improve the opportunity of such treatments. Transcription factors play important roles in regulating cell proliferation and differentiation. RUNX1 is a transcription factor that is expressed in blood cells and regulates the expression of many blood cell related genes. Therefore, the specific aim of our studies is to examine the effect of RUNX1 on blood cell formation, expansion, and differentiation from human embryonic stem cells and induced pluripotent stem cells. During the second year of funding period, we have performed hematopoietic cell differentiation using both human ESCs and iPSCs in the presence and absence of this transcription factor. Our results indicate that this factor promotes the production of blood stem cells and progenitors. We will make additional amount of this factor and further confirm our initial finding.

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