SBIR-STTR Award

It is believed that emergent stem cell-based therapies are destined to replace islet transplantation in the near future. The first wave of next-generation cell therapeutics for type 1 diabetes (T1D) will likely take the shape of pluripotent stem cell (PSC)-derived pancreatic progenitors (PPs) that mature into insulin-producing cells upon transplantation. However, for all its promise, the success of this approach hinges on the assumption that the microenvironment that leads to effective maturation in a mouse model (which is not even diabetic at the time of transplantation in published reports) will be the same i human patients with autoimmune diabetes. Other considerations, such as the safety of partially differentiated PSC-derived products and lag time to functional maturation, will also have to be addressed. The use of fully differentiated insulin- producing cells has always been the first choice, but the above approach has prevailed owing to the inability of current culture standards at yielding functional ß cells. In collaboration with our partners at the University of Miami, Ophysio, Inc. has successfully developed a platform to aid in the terminal in vitro differentiation of PPs of different origins (PSC and native PPs). This patented technology is based on the accurate targeting of physiological oxygenation throughout cell aggregates in culture - which conventional means of culture fail to achieve. Oxygen tension lies at the crossroads of key pancreatic differentiation pathways, and its evolution throughout development has been conclusively shown to drive cell fate. Here we seek to apply these principles to the terminal maturation of a novel sub-population of PPs that our collaborators have described in human non-endocrine pancreatic tissue (hNEPT), which comprises 98% of the pancreas and is routinely discarded after islet isolation. This sub- population, unequivocally identified through i vitro lineage-tracing techniques, is characterized by its responsiveness to the FDA-approved bone morphogenetic protein 7 (BMP-7). hNEPT exposure to BMP-7 results in the efficient (up to 33% in preliminary data) generation of endocrine cells that secrete insulin in response to glucose stimulation in vitro and in vivo at levels that, under conditions that will be further explored in this proposal, approximate those of human islets. BMP-7-responsive PPs from hNEPT represent a valid alternative to PSC for clinical applications, as this technology capitalizes on current clinical strategies (islet isolation and transplantation) for which there ar already well established networks; increased safety of adult stem cell products vs. PSC-derived ones; and ease of in vitro expansion using a single, FDA-approved agent that is already in clinical trials for unrelated conditions. Coupled with Ophysio's technology for enhanced in vitro maturation, this approach has rapid translational potential for the effective treatment of T1D.

Public Health Relevance Statement:


Public Health Relevance:
Progress in the efficient generation of insulin-producing cells is expected to result in new cell therapies for type 1 diabetes. Ophysio, Inc. has successfully developed a platform to aid in the differentiation of such cells. Our patented technology is based on the accurate provision of physiological oxygenation, which conventional means of culture cannot achieve. Here we seek to apply this principle to the maturation of a novel sub- population of pancreatic progenitor cells that our collaborators at the University of Miami have described in human non-endocrine pancreatic tissue (hNEPT), which comprises 98% of the pancreas and is routinely discarded after the insulin-producing islets (used for islet transplantation) are isolate. These progenitors are characterized by their responsiveness to BMP-7, an FDA-approved agent. Coupled with Ophysio's technology for enhanced maturation, this approach has rapid translational potential for the effective treatment of T1D, and as such is highly aligned with the NIH mission.

Project Terms:
Academia; Address; Administrator; adult stem cell; Autoimmune Diabetes; Award; base; Beta Cell; Biology; bone morphogenetic protein 7; Cell Differentiation process; Cell Maturation; Cell Therapy; Cells; Clinical; clinical application; Clinical Trials; Collaborations; Consensus; Coupled; Data; design; Development; Diabetes Mellitus; diabetic; Ductal; effective therapy; Endocrine; Evolution; experience; Exposure to; FDA approved; Future; Generations; Glucose; Goals; Growth; Hormones; Human; human embryonic stem cell; human tissue; In Vitro; in vivo; Industry; Insulin; Insulin-Dependent Diabetes Mellitus; Investigational New Drug Application; islet; Islets of Langerhans Transplantation; Knowledge; Laboratories; Legal patent; Length; Mature Bone; Methods; Mission; mouse model; Mus; Natural regeneration; next generation; non-diabetic; novel; Organ; Oxygen; Oxygen measurement, partial pressure, arterial; Pancreas; Pathway interactions; Patients; Phase; phase 2 study; Physiological; Pluripotent Stem Cells; Population; pre-clinical; prevent; Procedures; progenitor; public health relevance; Publishing; Reporting; Research; Research Institute; research study; response; Safety; Shapes; Small Business Innovation Research Grant; Source; Staging; stem cell differentiation; Stem cells; Streptozocin; Structure of beta Cell of islet; success; Techniques; Technology; technology development; Testing; Therapeutic; Time; Tissues; Translational Research; Transplantation; United States National Institutes of Health; Universities; Washington; Work

Islet transplantation represents the current cell therapy standard for type 1 diabetes (T1D). However, the gap between the availability of donor organs and the clinical demand for them calls for the development of alternative/renewable sources of insulin-producing cells. In addition to this therapeutic need, a steady supply of islets is also needed for research and drug discovery purposes. Human embryonic stem cells (hESc) differentiated into pancreatic -cell precursors are presently the subject of Phase I/II clinical trials. However, the success of this approach hinges on the assumption that the microenvironment that leads to effective maturation in a mouse model will be the same in human patients with autoimmune diabetes. The safety of partially differentiated hESc-derived products, efficacy of the macro- encapsulation devices used to shield them from allo- and auto-immunity, and lag time to functional maturation remain open questions. The use of insulin-producing cells that are mature and functional at the time of transplantation may circumvent some of these problems. However, despite claims to the contrary, there is no current protocol to date that yields -like cells capable of reversing diabetes right after transplantation. In collaboration with our partners at the University of Miami, Ophysio, Inc. has successfully developed a platform to aid in the terminal in vitro differentiation of pancreatic progenitors (PPs) of different origins (hESc and native murine pancreas). This patented technology is based on the accurate targeting of physiological oxygenation throughout cell aggregates in culture –which conventional means of culture fail to achieve. Oxygen tension lies at the crossroads of key pancreatic differentiation pathways, and its evolution throughout development has been conclusively shown to drive cell fate. Here we seek to extend these principles to the terminal maturation of a novel sub-population of PPs that our collaborators have described in human non-endocrine pancreatic tissue (hNEPT), which comprises 98% of the pancreas and is routinely discarded after islet isolation. This sub-population, identified through in vitro lineage-tracing techniques, is characterized by its responsiveness to the FDA- approved bone morphogenetic protein 7 (BMP-7). hNEPT exposure to BMP-7 results in the efficient (up to 15% in preliminary data) generation of endocrine cells that secrete insulin at levels that fall right within the range published for human isolated islets and exhibit robust glucose responsiveness in vitro and in vivo. Our Phase II studies aim at capitalizing on our Phase I data. These include not only the proof of principle that oxygen modulation improves BMP-7-mediated conversion of hNEPT, but also new findings on the phenotype of BMP-7 responsive cells that will allow for their prospective isolation from raw hNEPT preparations. Our specific aims are: (1) To determine whether in vitro targeting of physiological pO2 in PDX1 (P2RY1)+/ALK3+-sorted hNEPT subpopulations results in functional -like cells capable of reversing diabetes in mice; and (2) To scale up the process using an entire organ (10-12 ml of hNEPT pellet after islet isolation) using Ophysio’s new T75 oxygen-modulating devices (designed in the context of our previous award 2R44DK083832-02). In addition to the optimization and scale up of the process, we will simultaneously establish cGMP manufacturing protocols, file for IP protection of the final method and begin licensing contracts with parties for use of the process to obtain the cells for research purposes. We contend that BMP-7-responsive PPs from hNEPT represent a valid alternative to hESc for clinical applications, as this technology capitalizes on current clinical strategies (islet isolation and transplantation) for which there are already well established networks; increased safety of adult cell products vs. hESc-derived ones; and ease of in vitro expansion/differentiation using a single, FDA-approved clinical product. Coupled with Ophysio’s technology for enhanced in vitro maturation, this approach has rapid translational potential for the treatment of diabetes mellitus.

Public Health Relevance Statement:
NARRATIVE Progress in the efficient generation of insulin-producing cells is expected to result in new cell therapies for type 1 and 2 diabetes (T1/2D). Ophysio, Inc. has successfully developed a platform to aid in the differentiation of such cells from different sources. Our patented technology is based on the accurate provision of physiological oxygenation, which conventional means of culture cannot achieve. Here we seek to expand the application of this principle to the maturation of a novel sub-population of pancreatic progenitor cells that our collaborators at the University of Miami have described in human non-endocrine pancreatic tissue (hNEPT), which comprises 98% of the pancreas and is routinely discarded after the insulin-producing islets (used for islet transplantation) are isolated. These progenitors are characterized by their responsiveness to BMP-7, an FDA-approved agent. Coupled with Ophysio’s technology for enhanced maturation, this approach has rapid translational potential for the effective treatment of T1/2D, and as such is highly aligned with the NIH mission.

NIH Spending Category:
Autoimmune Disease; Biotechnology; Diabetes; Regenerative Medicine; Stem Cell Research; Stem Cell Research - Embryonic - Non-Human; Stem Cell Research - Nonembryonic - Human; Transplantation

Project Terms:
Academia; Administrator; Adult; Area; Autoimmune Diabetes; Automobile Driving; Award; base; Beta Cell; Biology; Biotechnology; BMP7 gene; Cell Lineage; Cell Maturation; Cell Therapy; cell type; Cells; Clinical; clinical application; Clinical Trials; Collaborations; Contracts; Coupled; Cyclic GMP; Data; design; Development; Device Designs; Devices; Diabetes Mellitus; drug discovery; effective therapy; Embryo; Endocrine; Evolution; Exhibits; experience; Exposure to; falls; FDA approved; Funding; Generations; Glucose; Growth; Human; human embryonic stem cell; human tissue; Immunity; improved; In Situ; In Vitro; in vivo; Industry; Insulin; Insulin-Dependent Diabetes Mellitus; islet; Islets of Langerhans Transplantation; Knowledge; Lead; Legal patent; Length; Licensing; Mediating; Methods; Mission; mouse model; Mus; Natural regeneration; Nature; Non-Insulin-Dependent Diabetes Mellitus; novel; Organ; Organ Donor; Oxygen; Pancreas; Pathway interactions; Patients; Phase; phase 2 study; Phenotype; Physiological; Pluripotent Stem Cells; Population; precursor cell; Preparation; Procedures; Process; Production; progenitor; Progress Reports; prospective; Protocols documentation; Publications; Publishing; receptor; Reporting; Research; Research Institute; research study; response; Safety; scale up; Sorting - Cell Movement; Source; Stem cells; success; Techniques; Technology; Testing; Therapeutic; Time; tissue preparation; Tissues; Traction; Transplantation; United States National Institutes of Health; Universities; Was