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| Content Provider | frontiers |
|---|---|
| Author | Kumar, Bijender Afshar-Kharghan, Vahid Mendt, Mayela Sackstein, Robert Tanner, Mark R. Popat, Uday Ramdial, Jeremy Daher, May Jimenez, Juan Basar, Rafet Melo Garcia, Luciana Shanley, Mayra Kaplan, Mecit Wan, Xinhai Nandivada, Vandana Reyes Silva, Francia Woods, Vernikka Gilbert, April Gonzalez-Delgado, Ricardo Acharya, Sunil Lin, Paul Rafei, Hind Banerjee, Pinaki Prosad Shpall, Elizabeth J. |
| Abstract | The current global platelet supply is often insufficient to meet all the transfusion needs of patients, in particular for those with alloimmune thrombocytopenia. To address this issue, we have developed a strategy employing a combination of approaches to achieve more efficient production of functional megakaryocytes (MKs) and platelets collected from cord blood (CB)-derived CD34+ hematopoietic cells. This strategy is based on ex-vivo expansion and differentiation of MKs in the presence of bone marrow niche-mimicking mesenchymal stem cells (MSCs), together with two other key components: (1) To enhance MK polyploidization, we used the potent pharmacological Rho-associated coiled-coil kinase (ROCK) inhibitor, KD045, resulting in liberation of increased numbers of functional platelets both in-vitro and in-vivo; (2) To evade HLA class I T-cell-driven killing of these expanded MKs, we employed CRISPR-Cas9-mediated beta-2 microglobulin (beta2M) gene knockout (KO). We found that coculturing with MSCs and MK-lineage-specific cytokines significantly increased MK expansion. This was further increased by ROCK inhibition, which induced MK polyploidization and platelet production. Additionally, ex-vivo treatment of MKs with KD045 resulted in significantly higher levels of engraftment and donor chimerism in a mouse model of thrombocytopenia. Finally, beta2M KO allowed MKs to evade killing by allogeneic T-cells. Overall, our approaches offer a novel, readily translatable roadmap for producing adult donor-independent platelet products for a variety of clinical indications. |
| ISSN | 16643224 |
| DOI | 10.3389/fimmu.2022.1018047 |
| Volume Number | 13 |
| Journal | Frontiers in Immunology |
| Language | English |
| Publisher Date | 2022-09-20 |
| Access Restriction | Open |
| Subject Keyword | Megakaryocyte Platelet Cord blood Thrombocytopaenia Beta2 microglobin Rho-associated coiled coil-containing protein kinase |
| Content Type | Text |
| Resource Type | Article |
| Subject | Immunology and Allergy Immunology |
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