Michael Kyba, Ph.D.
Cancer and Cardiovascular Research Building (CCRB)
2231 6th St SE
Minneapolis, MN 55455
Office 4-127 / Lab 4-240
Deriving therapeutic hematopoietic stem cells from embryonic stem cells.
ES cells are totipotent and capable of recapitulating all of the developmental events of embryogenesis. They are therefore theoretically the ideal source of cells for regenerative therapies. However, turning theory into practice is not straightforward, and very few successful models of such therapy exist. We have shown that in the case of the hematopoietic system, ES cells are programmed to undergo an embryonic mode of hematopoiesis, which produces a different array of cell types than the adult, and derives from a distinct, so called “primitive” hematopoietic stem cell. The primitive hematopoietic stem cell is incapable of engrafting when transplanted into lethally irradiated adult recipients. To derive adult, definitive hematopoietic stem cells, a developmental maturation process must be induced. Our goal is to understand the molecular details of this process: what secreted factors specify definitive hematopoiesis, and what key differences distinguish the primitive and definitive hematopoietic stem cells.
ES Cells Derived Blood ColoniesThe Hox code for hematopoietic stem cell self-renewal.
Gain of function studies with HoxB4 have shown that this Hox family member is involved in the regulation of self-renewal. Unfortunately, because other Hox genes cause leukemia when constitutively expressed, they have largely been ignored. By using conditional gene expression, we have shown that HoxB4 is neither unique in promoting hematopoietic stem cell self-renewal, nor most potent. We wish to understand how Hox genes control stem cell self-renewal, and are identifying regulatory circuits under Hox control.
Specification of Hematopoietic Mesoderm.
HoxB4 plays a role in the maturation of hematopoietic stem cells. We are also interested in factors that specify the origin of the hematopoietic stem cell in early mesoderm. Our ultimate goal is the synchronous generation of hematopoietic stem cells from ES cells. While therapeutically useful, it is also the gold standard by which we can judge our understanding of the process: if we can enforce with certainty a given lineage on the progeny of an ES cell, then we can be satisfied that we understand, at least at a basic level, the process of lineage selection.
Skeletal muscle stem cells and FSH muscular dystrophy.
Certain degenerative diseases may be the result of ineffective self-renewal or differentiation of lineage specific stem cells. We are particularly interested in Fascioscapulohumeral Muscular Dystrophy (FSHD), a dominant dystrophy associated with a contraction of 4q subtelomeric repeats. Although the condition is almost certainly caused by derepression of a gene in the viscinity of 4q, the protein products of candidate genes in this area can not be detected overexpressed in patient muscle samples. Because muscle stem cells (satellite cells) are rare, proteins overexpressed specifically in satellite cells are unlikely to be identified in patient biopsies. We are testing the hypothesis that a Hox gene embedded within the 4q repeats, DUX4, causes FSHD when derepressed in muscle satellite cells.
- Dandapat A, Bosnakovski D, Baltgalvis KA, Vang D, Hartweck L, Baik J, Nash N, Darabi R, PerlingeiroRCR, Hamra FK, Gupta K, Lowe DA, Kyba M, (2014) “Dominant lethal pathologies in male mice engineered to contain an X-linked DUX4 transgene.” Cell Reports 8:1-13.
- Bosnakovski D, Choi SH, Strasser JM, Toso E, Walters MA, Kyba M, (2014) “High-throughput screening identifies inhibitors of DUX4-induced myoblast toxicity” Skeletal Muscle 4(1):4.
- Iacovino M, Roth ME, Kyba M. (2014) “Rapid genetic modification of mouse embryonic stem cells by inducible cassette exchange.” Methods Mol. Biol. 1101:339-351.
- Chan SS, Shi X, Toyama A, Arpke RW, Dandapat A, Iacovino M, Kang JJ, Le G, Hagen HR, Garry DJ, Kyba M, (2013) “Mesp1 patterns mesoderm into cardiac, hematopoietic, or skeletal myogenic progenitors in a context-dependent manner” Cell Stem Cell 12:587-601.
- Arpke RW, Darabi R, Mader TL, Zhang Y, Toyama A, Lonetree CL, Nash N, Lowe DA, Perlingeiro RC, Kyba M, (2013) “A new immuno-dystrophin-deficient model, the NSG-mdx4Cv mouse, provides evidence for functional improvement following allogeneic satellite cell transplantation.” Stem Cells 19:e1402
- Dandapat A, Hartweck L, Kyba M, (2013) “Expression of the human FSHD-linked DUX4 gene induces neurogenesis during differentiation of murine embryonic stem cells.” Stem Cells and Development 22:2440-2448
- Filareto A, Parker S, Darabi R, Borges L, Iacovino M, Schaaf T, Mayerhofer T, Chamberlain JS, Ervasti JM, McIvor RS, Kyba M, Perlingeiro RC. (2013) "An ex vivo gene therapy approach to treat muscular dystrophy using inducible pluripotent stem cells." Nat Commun. 4:1549
- Kyba M. (2013) “Hemogenic endothelium in a dish” Blood 121:417-418.
- Hartweck LM, Anderson LJ, Lemmers RJ, Dandapat A, Toso EA, Dalton JC, Tawil R, Day JW, van der Maarel SM,Kyba M. (2013) “A focal domain of extreme demethylation within D4Z4 in FSHD2.” Neurology 80:392-399.
Older papers of significance to the field:
- Antonica F, Kasprzyk DF, Opitz R, Iacovino M, Liao XH, Dumitrescu AM, Refetoff S, Peremans K, Manto M, Kyba M, Costagliola S. (2012) “Generation of functional thyroid from embryonic stem cells” Nature 491:66-71.
- Darabi R, Arpke RW, Irion S, Dimos JT, Grskovic M, Kyba M, Perlingeiro RC (2012) “Human ES- and iPS-derived myogenic progenitors restore dystrophin and improve contractility upon transplantation in dystrophic mice” Cell Stem Cell 10:610-619.
- Bosnakovski D, Xu Z, Gang EJ, Galindo CL, Liu M, Simsek T, Garner HR, Agha-Mohammadi S, Tassin A, Frédérique Coppée, Belayew A, Perlingeiro RCR, Kyba M, (2008) “An isogenetic myoblast screen identifies DUX4-mediated FSHD-associated molecular pathologies” EMBO J. 27:2766-2779.
- Bosnakovski D, Xu Z, Li W, Thet S, Cleaver O, Perlingeiro RCR, Kyba M, (2008) “Prospective isolation of skeletal muscle stem cells with a Pax7 reporter” Stem Cells 26:3194-3204.
- Tang W, Zeve D, Suh J, Bosnakovski D, Kyba M, Hammer RE, Tallquist MD, Graff JM. (2008) “White fat progenitor cells reside in the Adipose Vasculature” Science 322:583-536.
- Darabi, R, Gelbach, K, Bachoo, RM, Kamath, S, Osawa, M, Kamm, KE, Kyba M, Perlingeiro, RCR. (2008) “Functional skeletal muscle regeneration from differentiating embryonic stem cells” Nature Medicine, 14:134-143.
- Kyba, M, Perlingeiro, RCR, Daley, GQ. HoxB4 Confers Definitive Lymphoid-Myeloid Engraftment Potential on Embryonic Stem Cell and Yolk Sac Hematopoietic Progenitors. (2002) Cell 109:29-37.