Functional Consequences of 17q21.31/WNT3-WNT9B Amplification in hPSCs with Respect to Neural Differentiation
Chun-Ting Lee,
Raphael M. Bendriem,
Abigail A. Kindberg,
Lila T. Worden,
Melanie P. Williams,
Tomas Drgon,
Barbara S. Mallon,
Brandon K. Harvey,
Christopher T. Richie,
Rebecca S. Hamilton,
Jia Chen,
Stacie L. Errico,
Shang-Yi A. Tsai,
George R. Uhl,
William J. Freed
Affiliations
Chun-Ting Lee
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Raphael M. Bendriem
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Abigail A. Kindberg
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Lila T. Worden
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Melanie P. Williams
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Tomas Drgon
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Barbara S. Mallon
NIH Stem Cell Unit, Intramural Research Program, National Institute of Neurological Disorders and Stroke, Department of Health and Human Services, NIH, Bethesda, MD 20892, USA
Brandon K. Harvey
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Christopher T. Richie
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Rebecca S. Hamilton
NIH Stem Cell Unit, Intramural Research Program, National Institute of Neurological Disorders and Stroke, Department of Health and Human Services, NIH, Bethesda, MD 20892, USA
Jia Chen
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Stacie L. Errico
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Shang-Yi A. Tsai
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
George R. Uhl
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
William J. Freed
Intramural Research Program, National Institute on Drug Abuse, Department of Health and Human Services, NIH, Baltimore, MD 21224, USA
Human pluripotent stem cell (hPSC) lines exhibit repeated patterns of genetic variation, which can alter in vitro properties as well as suitability for clinical use. We examined associations between copy-number variations (CNVs) on chromosome 17 and hPSC mesodiencephalic dopaminergic (mDA) differentiation. Among 24 hPSC lines, two karyotypically normal lines, BG03 and CT3, and BG01V2, with trisomy 17, exhibited amplification of the WNT3/WNT9B region and rapid mDA differentiation. In hPSC lines with amplified WNT3/WNT9B, basic fibroblast growth factor (bFGF) signaling through mitogen-activated protein kinase (MAPK)/ERK amplifies canonical WNT signaling by phosphorylating LRP6, resulting in enhanced undifferentiated proliferation. When bFGF is absent, noncanonical WNT signaling becomes dominant due to upregulation of SIAH2, enhancing JNK signaling and promoting loss of pluripotency. When bFGF is present during mDA differentiation, stabilization of canonical WNT signaling causes upregulation of LMX1A and mDA induction. Therefore, CNVs in 17q21.31, a “hot spot” for genetic variation, have multiple and complex effects on hPSC cellular phenotype.
