Production of red blood cells and platelets from stem cells
Inventors
Murphy, George J. • Sherr, David H. • Rozelle, Sarah S. • Smith, Brenden W.
Assignees
Boston University • Boston Medical Center Corp
Publication Number
US-12173320-B2
Publication Date
2024-12-24
Expiration Date
2033-03-14
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
This disclosure provides methods of making a megakaryocyte-erythroid progenitor cell (MEP), comprising differentiating a MEP precursor cell into a MEP in culture in the presence of an aryl hydrocarbon receptor (AhR) modulator. In some embodiments the AhR modulator is an AhR antagonist. In some embodiments the AhR modulator is an AhR agonist. In some embodiments the methods comprise culturing MEP precursor cells in the presence of an AHR antagonist and then culturing MEP precursor cells in the presence of an AHR agonist. In some embodiments the stem cell is a pluripotent stem cell. In some embodiments the MEP co-expresses CD41 and CD235. In some embodiments the number of MEPs produced in the culture increases exponentially. Methods of making a red blood cell (RBC) by culturing a MEP in the presence of an AhR modulator are also provided. Methods of making a megakaryocyte and/or a platelet, comprising culturing a MEP in the presence of an AhR modulator are also provided. In some embodiments the AhR modulator is an AhR antagonist. This disclosure also provides compositions comprising at least 1 million MEPs per ml and compositions in which at least 50% of the cells are MEPs, among other things.
Core Innovation
This invention provides methods for making megakaryocyte-erythroid progenitor cells (MEPs), red blood cells (RBCs), megakaryocytes (Mks), and platelets by differentiating precursor cells in culture using aryl hydrocarbon receptor (AhR) modulators. Specifically, the methods utilize AhR antagonists and/or agonists at defined steps to direct lineage specification and expansion. The invention covers the use of both pluripotent stem cells (such as iPSCs or ESCs) and other hematopoietic precursors as starting materials.
A problem addressed by this invention is the lack of efficient, robust, and scalable platforms for producing sufficient quantities of blood products, particularly RBCs and platelets, for transfusion and research purposes. Conventional methods often result in limited cell numbers, require complex or undefined culture conditions, or fail to generate cell populations with the requisite purity or scalability.
The disclosed methods achieve exponential expansion of MEPs in feeder-free, chemically defined systems by modulating AhR signaling, especially using potent AhR agonists such as FICZ. Furthermore, sequential use of AhR agonists and antagonists enables precise control over differentiation into either erythroid or megakaryocytic lineages, allowing for the efficient in vitro production of functional RBCs and platelets from patient- or donor-specific sources. The patent also describes compositions enriched for these cells and their use in therapeutic contexts.
Claims Coverage
There are two independent claims, each covering specific inventive features regarding the use of AhR modulation in differentiating MEPs toward megakaryocytes.
Culturing MEPs with sequential AhR agonist and antagonist exposure to produce megakaryocytes
The process involves obtaining a population of megakaryocyte-erythroid progenitor cells (MEP) that have been differentiated from MEP precursor cells by culturing in the presence of an aryl hydrocarbon receptor (AhR) antagonist. This population of MEP cells is then cultured in the presence of an AhR agonist prior to culturing the MEP cells in the presence of an AhR antagonist to make a megakaryocyte (Mk).
Obtaining MEPs via AhR agonist differentiation and sequential exposure to AhR antagonist and agonist to make megakaryocytes
The method comprises obtaining MEP cells by differentiating MEP precursor cells into MEPs in culture in the presence of an AhR agonist. The MEPs are then cultured in the presence of an AhR antagonist and subsequently in the presence of an AhR agonist to produce megakaryocytes.
The independent claims focus on specific sequences of AhR modulation—using agonists and antagonists at defined stages—to precisely direct the differentiation of MEPs into megakaryocytes. The inventive features are the sequential application and manipulation of AhR modulators to control hematopoietic differentiation.
Stated Advantages
The methods enable exponential and robust expansion of MEPs and efficient production of RBCs and platelets in chemically defined, feeder-free culture conditions.
The invention provides a scalable platform for producing large quantities of blood cell types required for transfusion therapy and research.
The protocols are serum-free and feeder-free, reducing the risk of contamination and thus enhancing the clinical safety of derived cells.
Patient-specific or matched genetic identity of derived cells can be achieved using iPSC sources, potentially minimizing immunogenicity in transfusions.
Precise modulation of differentiation steps enables high-purity populations of targeted blood cell types, suitable for therapeutic or screening applications.
Documented Applications
Production of RBCs and platelets for transfusion therapy in patients with anemia, including sickle cell anemia and thalassemia, or thrombocytopenia.
Preparation of compositions containing enriched populations of RBCs, MEPs, megakaryocytes, and/or platelets for clinical use.
Methods of treating anemia and thrombocytopenia in patients by administering transfusion compositions derived using these methods.
Screening compounds for effects on RBCs, Mks, or platelets using in vitro differentiated cells for research and drug discovery.
Increasing the platelet count of a mammal by administering an AhR agonist.
Interested in licensing this patent?