TY - JOUR
T1 - Splicing factor deficits render hematopoietic stem and progenitor cells sensitive to STAT3 inhibition
AU - Potts, Kathryn S.
AU - Cameron, Rosannah C.
AU - Metidji, Amina
AU - Ghazale, Noura
AU - Wallace, La Shanale
AU - Leal-Cervantes, Ana I.
AU - Palumbo, Reid
AU - Barajas, Juan Martin
AU - Gupta, Varun
AU - Aluri, Srinivas
AU - Pradhan, Kith
AU - Myers, Jacquelyn A.
AU - McKinstry, Mia
AU - Bai, Xiaoying
AU - Choudhary, Gaurav S.
AU - Shastri, Aditi
AU - Verma, Amit
AU - Obeng, Esther A.
AU - Bowman, Teresa V.
N1 - Funding Information:
T.V.B. was funded by Gabrielle's Angel Foundation , American Cancer Society RSG-129527-DDC , Kimmel Foundation , the Edward P. Evans Foundation , DOD BM180109 , NIH / NIDDK 1R56DK121738 , and 1R01DK121738 ; K.S.P. was funded the American Australian Association Sir Rupert Murdoch Postdoctoral Fellowship and The Einstein Training Program in Stem Cell Research funded by the Empire State Stem Cell Fund through New York State Department of Health Contract C30292GG ; R.C.C. was funded by the Aplastic Anemia and MDS International Foundation Fellowship; N.G. was funded by T32GM007491 ; E.A.O. was funded by the Edward P. Evans Foundation , the American Society of Hematology , and Gabrielle's Angel Foundation ; and V.G. was supported by 5R01GM057829 funded to Charles Query.
Funding Information:
We would like to thank H3 Biomedicine for providing E7107 reagent. We thank Jackie Boultwood and Andrea Pellagatti for sharing their MDS gene expression and mutational microarray dataset. We thank Eirini Trompouki, Meelad Dawlaty, and Charles Query for helpful discussions on this work, Scott Cameron for advice and assistance on computational aspects of the work, and Anastasia Nizhnik and Ilana Karp for technical assistance. We also want to acknowledge the assistance of numerous core facilities at Albert Einstein College of Medicine, including Flow Cytometry and Genomics Facilities (funded by NCI Cancer grant P30CA013330 ), and the Zebrafish Core Facility. We would like to thank Shondra Miller and the Center for Advanced Genome Engineering at St. Jude Children’s Research Hospital for generating the mutant K562 cells and the Vanderbilt University School of Medicine Technologies for Advanced Genomics scientific core facility for RNA sequencing. The graphical abstract was made in BioRender.
Funding Information:
T.V.B. was funded by Gabrielle's Angel Foundation, American Cancer Society RSG-129527-DDC, Kimmel Foundation, the Edward P. Evans Foundation, DOD BM180109, NIH/NIDDK 1R56DK121738, and 1R01DK121738; K.S.P. was funded the American Australian Association Sir Rupert Murdoch Postdoctoral Fellowship and The Einstein Training Program in Stem Cell Research funded by the Empire State Stem Cell Fund through New York State Department of Health Contract C30292GG; R.C.C. was funded by the Aplastic Anemia and MDS International Foundation Fellowship; N.G. was funded by T32GM007491; E.A.O. was funded by the Edward P. Evans Foundation, the American Society of Hematology, and Gabrielle's Angel Foundation; and V.G. was supported by 5R01GM057829 funded to Charles Query. We would like to thank H3 Biomedicine for providing E7107 reagent. We thank Jackie Boultwood and Andrea Pellagatti for sharing their MDS gene expression and mutational microarray dataset. We thank Eirini Trompouki, Meelad Dawlaty, and Charles Query for helpful discussions on this work, Scott Cameron for advice and assistance on computational aspects of the work, and Anastasia Nizhnik and Ilana Karp for technical assistance. We also want to acknowledge the assistance of numerous core facilities at Albert Einstein College of Medicine, including Flow Cytometry and Genomics Facilities (funded by NCI Cancer grant P30CA013330), and the Zebrafish Core Facility. We would like to thank Shondra Miller and the Center for Advanced Genome Engineering at St. Jude Children's Research Hospital for generating the mutant K562 cells and the Vanderbilt University School of Medicine Technologies for Advanced Genomics scientific core facility for RNA sequencing. The graphical abstract was made in BioRender. K.S.P. R.C.C. E.A.O. and T.V.B. designed the project experimental approach. K.S.P. R.C.C. N.G. L.W. A.M. A.I.L.-C. R.P. J.M.B. and M.M. performed the experiments and analyzed the data. V.G. K.P. and J.A.M. provided bioinformatics support. S.A. X.B. G.S.C. A.S. and A.V. provided reagents, critical advice, and patient samples. J.M.B. L.W. A.I.L.-C. A.M. R.P. and E.A.O. provided engineered K562 cells and performed mouse experiments. K.S.P. and T.V.B. wrote the manuscript. All authors reviewed and approved the manuscript. A.V. participates on the advisory boards for Stelexis, Bakx, Novartis, Acceleron, and Celgene, which includes stock options, received a consultation fee from Janssen Pharmaceuticals, and received research funding from Prelude, BMS, GSK, Incyte, Medpacto, Curis, and Eli Lilly. A.S. has received an honorarium from GLG, Guidepoint, Onclive, a consultation fee from Janssen Pharmaceuticals, an advisory board fee from Rigel Pharmaceuticals, and research funding from Kymera Therapeutics.
Funding Information:
A.V. participates on the advisory boards for Stelexis, Bakx, Novartis, Acceleron, and Celgene, which includes stock options, received a consultation fee from Janssen Pharmaceuticals, and received research funding from Prelude, BMS, GSK, Incyte, Medpacto, Curis, and Eli Lilly. A.S. has received an honorarium from GLG, Guidepoint, Onclive, a consultation fee from Janssen Pharmaceuticals, an advisory board fee from Rigel Pharmaceuticals, and research funding from Kymera Therapeutics.
Publisher Copyright:
© 2022 The Author(s)
PY - 2022/12/13
Y1 - 2022/12/13
N2 - Hematopoietic stem and progenitor cells (HSPCs) sustain lifelong hematopoiesis. Mutations of pre-mRNA splicing machinery, especially splicing factor 3b, subunit 1 (SF3B1), are early lesions found in malignancies arising from HSPC dysfunction. However, why splicing factor deficits contribute to HSPC defects remains incompletely understood. Using zebrafish, we show that HSPC formation in sf3b1 homozygous mutants is dependent on STAT3 activation. Clinically, mutations in SF3B1 are heterozygous; thus, we explored if targeting STAT3 could be a vulnerability in these cells. We show that SF3B1 heterozygosity confers heightened sensitivity to STAT3 inhibition in zebrafish, mouse, and human HSPCs. Cells carrying mutations in other splicing factors or treated with splicing modulators are also more sensitive to STAT3 inhibition. Mechanistically, we illustrate that STAT3 inhibition exacerbates aberrant splicing in SF3B1 mutant cells. Our findings reveal a conserved vulnerability of splicing factor mutant HSPCs that could allow for their selective targeting in hematologic malignancies.
AB - Hematopoietic stem and progenitor cells (HSPCs) sustain lifelong hematopoiesis. Mutations of pre-mRNA splicing machinery, especially splicing factor 3b, subunit 1 (SF3B1), are early lesions found in malignancies arising from HSPC dysfunction. However, why splicing factor deficits contribute to HSPC defects remains incompletely understood. Using zebrafish, we show that HSPC formation in sf3b1 homozygous mutants is dependent on STAT3 activation. Clinically, mutations in SF3B1 are heterozygous; thus, we explored if targeting STAT3 could be a vulnerability in these cells. We show that SF3B1 heterozygosity confers heightened sensitivity to STAT3 inhibition in zebrafish, mouse, and human HSPCs. Cells carrying mutations in other splicing factors or treated with splicing modulators are also more sensitive to STAT3 inhibition. Mechanistically, we illustrate that STAT3 inhibition exacerbates aberrant splicing in SF3B1 mutant cells. Our findings reveal a conserved vulnerability of splicing factor mutant HSPCs that could allow for their selective targeting in hematologic malignancies.
KW - CP: Molecular biology
KW - CP: Stem cell research
KW - SF3B1
KW - STAT3
KW - hematopoietic stem and progenitor cell
KW - myelodysplastic syndrome
KW - splicing factor
KW - zebrafish
UR - http://www.scopus.com/inward/record.url?scp=85144588190&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85144588190&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2022.111825
DO - 10.1016/j.celrep.2022.111825
M3 - Article
C2 - 36516770
AN - SCOPUS:85144588190
SN - 2211-1247
VL - 41
JO - Cell Reports
JF - Cell Reports
IS - 11
M1 - 111825
ER -