(1) 4D Bioprinting of Hematopoietic Tissues for Cellular Therapies
HSC transplantation (HSCT) is used to treat patients with genetic blood disorders, bone marrow failure syndromes, leukemia, and lymphoma. Despite advances in umbilical-cord and haplo-identical HSCT, its therapeutic use is restricted due to difficulty in finding HLA-matched donors, particularly for children born to multi-ethnic parents. Even if one finds a suitable match, immunologic complications such as graft-versus-host disease, donor rejection, and high treatment-related mortality can compromise patient survival. However, these complications are eliminated by autologous transplant. Therefore, there is a critical need to identify new methods and sources to generate compatible HSCs from patient-derived tissues.
Our laboratory analyzes how blood vessels give rise to hematopoietic stem cells (HSC) and investigates factors regulating this process. During fetal development, a subset of endothelial cells in the aorta-gonad-mesonephros, termed hemogenic endothelial cells, change their fate to become HSCs. However, the identity and molecular nature of hemogenic endothelial cells are enigmatic. We are investigating new cell-intrinsic, cell-extrinsic, and pharmacological targets that could stimulate endothelial emergence of HSCs.
We are also investigating 5D microenvironment conditions that stimulate HSC formation, development, and differentiation into adult blood lineages. We use state-of-the-art organ-in-a-dish, bio-inspired micro-electric-mechanical system (MEMS) fabrication, 3D-bioprinting, bio-inspired matrices, 3D organoid development, gene-editing, microfluidics, cell-engineering, nanotechnology, machine learning, as well as computational approaches to develop scalable platform for cGMP-grade cellular therapies in a dish.
Du W, Hong S, Scapin G, Goulard M, Shah DI (2019). Directed Collective Cell Migration using 3D Bioprinted Micropatterns on Thermo-Responsive Surfaces for Myotube Formation. ACS Biomaterials Sciences & Engineering. In Press
Anderson HA*, Patch T*, Reddy PN, Hagedorn E, Kim PG, Soltis KA, Chen MJ, Tamplin OJ, Frye M, MacLean G, Huebner K, Bauer DE, Kanki JP, Vogin G, Huston N, Nguyen M, Fujiwara Y, Paw BH, Vestweber D, Zon LI, Orkin SH, Daley GQ, Shah DI (2015). Hematopoietic stem cells develop in the absence of endothelial cadherin 5 expression. Blood 126: 2811-2820. * Co-First Authors. Featured as a covery story.
Hagedorn EJ, Cillis JL, Curley CR, Patch TC, Li B, Blaser BW, Riquelme R, Zon LI, Shah DI (2016). Generation of parabiotic zebrafish embryos by surgical fusion of developing blastula. Journal of Visualized Experiments
Cooney JD, Hildick-Smith GJ, Shafizadeh E, McBride PF, Carroll KJ, Anderson H, Shaw GC, Tamplin OJ, Branco DS, Dalton AJ, Shah DI, Wong C, Gallagher PG, Zon LI, North TE, Paw BH (2012). Zebrafish growth factor independence transcription factors offer a comparative paradigm for regulating primitive and definitive hematopoietic lineages. Dev. Biol. 373:431-441.
Mouse ex vivo Incubation
a.k.a. Mouse Neonatal Intensive Care Unit (NICU)
Zebrafish Parabiotic Surgery
(2) Analysis of Genetic Blood Disorders
The decades of research to find a cure for genetic blood disorders have generated a platform for understating congenital anemias. Randomized mutagenesis in zebrafish and mouse have led to the identification of novel functionally important genes related to human hematological diseases. Examples include mitoferrin in variant human erythropoietic protoporphyria (EPP), transcription inhibitory factor-1 in erythroid lineage commitment, ferroportin in type 4 familial hemochromatosis, gluteradoxin-5 in hypochromic anemia, 5-aminolevulinic acid synthase (Alas2) in erythropoietic protoporphyria, Hemojuvelin and Hfe in hematochromasis, Sec15L1 in hemoglobin disease, and Steap3 in iron-deficiency anemia. The mechanisms regulating HSC development and differentiation, red cell maturation, as well as iron and hemoglobin homeostasis are, however, largely unknown.
To advance our understanding and treatment of congenital hematological diseases, our laboratory analyzes new genes and mechanisms regulating red blood cell and HSC development.
Shah DI, Takahashi-Makise N, Cooney JD, Li L, Schultz IJ, Pierce EL, Narla A, Seguin A, Hattangadi SM, Medlock A, Langer NB, Dailey TA, Hurst SN, Faccenda D, Wiwczar J, Heggers SK, Vogin G, Chen W, Chen C, Campagna DR, Brugnara C, Zhou Y, Ebert BL, Danial NN, Fleming MD, Ward DM, Campanella M, Dailey HA, Kaplan J, Paw BH. (2012) Mitochondrial Atpif1 regulates heme synthesis in developing erythroblasts. Nature 491: 608-612.
Chen C, Garcia-Santos D, Li L, Ishikawa Y, Seguin A, Shah DI, Li L, Hattangai SM, Fegan KH, Hildick-Smith GJ, Cooney JD, Chen W, King MJ, Schultz IJ, Yien Y, Dalton AJ, Kingsley PD, Palis J, Lodish HF, Ward DM, Kaplan J, Maeda T, Ponka P, Paw BH. (2013) Snx3 regulates recycling of the trasnferrin receptor and iron assimilation. Cell Metabolism 17: 343-352.
Hildick-Smith GJ, Cooney JD, Garone C, Kremer LS, Haack TB, Thon JN, Miyata N, Lieber DS, Calvo SE, Akman HO, Yien YY, Huston NC, Branco DS, Shah DI, Freedman ML, Koehler CM, Italiano JE Jr, Merkenschlager A, Beblo S, Strom TM, Meitinger T, Freisinger P, Donati MA, Prokisch H, Mootha VK, DiMauro S, Paw BH (2013). Macrocytic anemia and mitochondriopathy resulting from a defect in sideroflexin 4. Am J Hum Genet. 93:906-914.
Yien YY, Robledo RF, Schultz IJ, Takahashi-Makise N, Gwynn B, Bauer DE, Dass A, Yi G, Li L, Hildick-Smith GJ, Cooney JD, Pierce EL, Mohler K, Dailey TA, Miyata N, Kingsley PD, Garone C, Hattangadi SM, Huang H, Chen W, Keenan EM, Shah DI, Schlaeger TM, DiMauro S, Orkin SH, Cantor AB, Palis J, Koehler CM, Lodish HF, Kaplan J, Ward DM, Dailey HA, Phillips JD, Peters LL, Paw BH (2014). TMEM14C is required for erythroid mitochondrial heme metabolism. J Clin Invest. 124:4294-304.