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  • 1.
    Guo, Jingtao
    et al.
    Univ Utah, Sch Med, Dept Oncol Sci, Howard Hughes Med Inst, Salt Lake City, UT 84112 USA.;Univ Utah, Sch Med, Huntsman Canc Inst, Salt Lake City, UT 84112 USA..
    Grow, Edward J.
    Univ Utah, Sch Med, Dept Oncol Sci, Howard Hughes Med Inst, Salt Lake City, UT 84112 USA.;Univ Utah, Sch Med, Huntsman Canc Inst, Salt Lake City, UT 84112 USA..
    Yi, Chongil
    Univ Utah, Sch Med, Dept Oncol Sci, Howard Hughes Med Inst, Salt Lake City, UT 84112 USA.;Univ Utah, Sch Med, Huntsman Canc Inst, Salt Lake City, UT 84112 USA..
    Mlcochova, Hana
    Univ Oxford, Radcliffe Dept Med, MRC Weatherall Inst Mol Med, Oxford OX3 9DS, England..
    Maher, Geoffrey J.
    Univ Oxford, Radcliffe Dept Med, MRC Weatherall Inst Mol Med, Oxford OX3 9DS, England..
    Lindskog, Cecilia
    Uppsala University, Science for Life Laboratory, SciLifeLab. Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology.
    Murphy, Patrick J.
    Univ Utah, Sch Med, Dept Oncol Sci, Howard Hughes Med Inst, Salt Lake City, UT 84112 USA.;Univ Utah, Sch Med, Huntsman Canc Inst, Salt Lake City, UT 84112 USA..
    Wike, Candice L.
    Univ Utah, Sch Med, Dept Oncol Sci, Howard Hughes Med Inst, Salt Lake City, UT 84112 USA.;Univ Utah, Sch Med, Huntsman Canc Inst, Salt Lake City, UT 84112 USA..
    Carrell, Douglas T.
    Univ Utah, Hlth Sci Ctr, Ctr Reconstruct Urol & Mens Hlth, Dept Surg Androl Urol, Salt Lake City, UT 84122 USA..
    Goriely, Anne
    Univ Oxford, Radcliffe Dept Med, MRC Weatherall Inst Mol Med, Oxford OX3 9DS, England..
    Hotaling, James M.
    Univ Utah, Hlth Sci Ctr, Ctr Reconstruct Urol & Mens Hlth, Dept Surg Androl Urol, Salt Lake City, UT 84122 USA..
    Cairns, Bradley R.
    Univ Utah, Sch Med, Dept Oncol Sci, Howard Hughes Med Inst, Salt Lake City, UT 84112 USA.;Univ Utah, Sch Med, Huntsman Canc Inst, Salt Lake City, UT 84112 USA..
    Chromatin and Single-Cell RNA- Seq Profiling Reveal Dynamic Signaling and Metabolic Transitions during Human Spermatogonial Stem Cell Development2017In: Cell Stem Cell, ISSN 1934-5909, E-ISSN 1875-9777, Vol. 21, no 4, p. 533-546Article in journal (Refereed)
    Abstract [en]

    Human adult spermatogonial stem cells (hSSCs) must balance self-renewal and differentiation. To understand how this is achieved, we profiled DNA methylation and open chromatin (ATAC-seq) in SSEA4(+) hSSCs, analyzed bulk and single-cell RNA transcriptomes (RNA-seq) in SSEA4+ hSSCs and differentiating c-KIT+ spermatogonia, and performed validation studies via immunofluorescence. First, DNA hypomethylation at embryonic developmental genes supports their epigenetic "poising'' in hSSCs for future/embryonic expression, while core pluripotency genes (OCT4 and NANOG) were transcriptionally and epigenetically repressed. Interestingly, open chromatin in hSSCs was strikingly enriched in binding sites for pioneer factors (NFYA/B, DMRT1, and hormone receptors). Remarkably, single-cell RNA-seq clustering analysis identified four cellular/developmental states during hSSC differentiation, involving major transitions in cell-cycle and transcriptional regulators, splicing and signaling factors, and glucose/mitochondria regulators. Overall, our results outline the dynamic chromatin/transcription landscape operating in hSSCs and identify crucial molecular pathways that accompany the transition from quiescence to proliferation and differentiation.

  • 2.
    Kee, Nigel
    et al.
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden.;Karolinska Inst, Dept Cell & Mol Biol, S-17177 Stockholm, Sweden..
    Volakakis, Nikolaos
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden..
    Kirkeby, Agnete
    Lund Univ, Dept Expt Med Sci, Lund Stem Cell Ctr, S-22184 Lund, Sweden..
    Dahl, Lina
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden..
    Storvall, Helena
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden..
    Nolbrant, Sara
    Lund Univ, Dept Expt Med Sci, Lund Stem Cell Ctr, S-22184 Lund, Sweden..
    Lahti, Laura
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden..
    Björklund, Åsa K.
    Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Cell and Molecular Biology, Molecular Evolution. Uppsala University, Science for Life Laboratory, SciLifeLab.
    Gillberg, Linda
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden..
    Joodmardi, Eliza
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden..
    Sandberg, Rickard
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden.;Karolinska Inst, Dept Cell & Mol Biol, S-17177 Stockholm, Sweden..
    Parmar, Malin
    Lund Univ, Dept Expt Med Sci, Lund Stem Cell Ctr, S-22184 Lund, Sweden..
    Perlmann, Thomas
    Ludwig Inst Canc Res, Box 240, S-17177 Stockholm, Sweden.;Karolinska Inst, Dept Cell & Mol Biol, S-17177 Stockholm, Sweden..
    Single-Cell Analysis Reveals a Close Relationship between Differentiating Dopamine and Subthalamic Nucleus Neuronal Lineages2017In: Cell Stem Cell, ISSN 1934-5909, E-ISSN 1875-9777, Vol. 20, no 1, p. 29-40Article in journal (Refereed)
    Abstract [en]

    Stem cell engineering and grafting of mesencephalic dopamine (mesDA) neurons is a promising strategy for brain repair in Parkinson's disease (PD). Refinement of differentiation protocols to optimize this approach will require deeper understanding of mesDA neuron development. Here, we studied this process using transcriptome-wide single-cell RNA sequencing of mouse neural progenitors expressing the mesDA neuron determinant Lmx1a. This approach resolved the differentiation of mesDA and neighboring neuronal lineages and revealed a remarkably close relationship between developing mesDA and subthalamic nucleus (STN) neurons, while also highlighting a distinct transcription factor set that can distinguish between them. While previous hESC mesDA differentiation protocols have relied on markers that are shared between the two lineages, we found that application of these highlighted markers can help to refine current stem cell engineering protocols, increasing the proportion of appropriately patterned mesDA progenitors. Our results, therefore, have important implications for cell replacement therapy in PD.

  • 3.
    Rivera-Gonzalez, Guillermo C.
    et al.
    Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA..
    Shook, Brett A.
    Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA..
    Andrae, Johanna
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Neuroscience, Molecular Cell Biology.
    Holtrup, Brandon
    Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA..
    Bollag, Katherine
    Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA..
    Betsholtz, Christer
    Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Vascular Biology.
    Rodeheffer, Matthew S.
    Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA.;Yale Univ, Sect Comparat Med, New Haven, CT 06520 USA..
    Horsley, Valerie
    Yale Univ, Dept Mol Cellular & Dev Biol, New Haven, CT 06520 USA.;Yale Univ, Dept Dermatol, Yale Sch Med, New Haven, CT 06520 USA..
    Skin Adipocyte Stem Cell Self-Renewal Is Regulated by a PDGFA/AKT-Signaling Axis2016In: Cell Stem Cell, ISSN 1934-5909, E-ISSN 1875-9777, Vol. 19, no 6, p. 738-751Article in journal (Refereed)
    Abstract [en]

    Tissue growth and maintenance requires stem cell populations that self-renew, proliferate, and differentiate. Maintenance of white adipose tissue (WAT) requires the proliferation and differentiation of adipocyte stem cells (ASCs) to form postmitotic, lipid-filled mature adipocytes. Here we use the dynamic adipogenic program that occurs during hair growth to uncover an unrecognized regulator of ASC self-renewal and proliferation, PDGFA, which activates AKT signaling to drive and maintain the adipogenic program in the skin. Pdgfa expression is reduced in aged ASCs and is required for ASC proliferation and maintenance in the dermis, but not in other WATs. Our molecular and genetic studies uncover PI3K/AKT2 as a direct PDGFA target that is activated in ASCs during WAT hyperplasia and is functionally required for dermal ASC proliferation. Our data therefore reveal active mechanisms that regulate ASC self-renewal in the skin and show that distinct regulatory mechanisms operate in different WAT depots.

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