ABCG2-Expressing Clonal Repopulating Endothelial Cells Serve to Form and Maintain Blood Vessels

Yang Lin; Chang-Hyun Gil; Kimihiko Banno; Masataka Yokoyama; Matthew Wingo; Ellen Go; Nutan Prasain; Ying Liu; Takashi Hato; Hisamichi Naito; Taku Wakabayashi; Musia Sominskaia; Meng Gao; Kevin Chen; Fuqiang Geng; Jesus Maria Gomez Salinero; Sisi Chen; W Christopher Shelley; Momoko Yoshimoto; Sergio Li Calzi; Michael P Murphy; Kyoji Horie; Maria B Grant; Ryan Schreiner; David Redmond; David P Basile; Shahin Rafii; Mervin C Yoder

doi:10.1161/CIRCULATIONAHA.122.061833

ABCG2-Expressing Clonal Repopulating Endothelial Cells Serve to Form and Maintain Blood Vessels

Circulation. 2024 Apr 29. doi: 10.1161/CIRCULATIONAHA.122.061833. Online ahead of print.

Authors

Yang Lin^#^{1

2

3}, Chang-Hyun Gil^#^{2

4}, Kimihiko Banno^#^{2

5}, Masataka Yokoyama^{1

6}, Matthew Wingo⁷, Ellen Go^{2

8}, Nutan Prasain², Ying Liu¹, Takashi Hato⁹, Hisamichi Naito¹⁰, Taku Wakabayashi^{10

11

12}, Musia Sominskaia¹, Meng Gao¹, Kevin Chen¹, Fuqiang Geng¹, Jesus Maria Gomez Salinero¹, Sisi Chen^{2

3

13}, W Christopher Shelley², Momoko Yoshimoto², Sergio Li Calzi^{14

15}, Michael P Murphy⁴, Kyoji Horie⁵, Maria B Grant^{14

15}, Ryan Schreiner¹, David Redmond¹, David P Basile¹⁶, Shahin Rafii¹, Mervin C Yoder^{2

3

17}

Affiliations

¹ Division of Regenerative Medicine, Hartman Institute for Therapeutic Organ Regeneration, Ansary Stem Cell Institute, Department of Medicine, Weill Cornell Medicine, New York, NY. (Y. Lin, M. Yokoyama, Y. Liu, M.S., M.G., K.C., F.G., J.M.G.S., R.S., D.R., S.R.).
² Department of Pediatrics, Herman B. Wells Center for Pediatric Research (Y. Lin, C.-H.G., K.B., E.G., N.P., S.C., W.C.S., M. Yoshimoto, M.C.Y.).
³ Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis. (Y. Lin, S.C., M.C.Y.).
⁴ Department of Surgery, Indiana University School of Medicine, Indianapolis. (C.-H.G., M.P.M.).
⁵ Department of Physiology II, Nara Medical University, Kashihara, Nara, Japan (K.B., K.H.).
⁶ Department of Molecular Diagnosis, Graduate School of Medicine, Chiba University, Japan (M. Yokoyama).
⁷ Department of Cardiothoracic Surgery, Weill Cornell Medicine, New York, NY. (M.W.).
⁸ Department of Pediatrics, Division of Pediatric Rheumatology, Riley Hospital for Children, Indianapolis, IN (E.G.).
⁹ Department of Medicine, Indiana University School of Medicine, Indianapolis. (T.H.).
¹⁰ Department of Vascular Physiology, Kanazawa University School of Medicine, Japan (H.N., T.W.).
¹¹ Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Japan (T.W.).
¹² Mid Atlantic Retina, Wills Eye Hospital, Thomas Jefferson University, Philadelphia, PA (T.W.).
¹³ Human Oncology and Pathogenesis Program, Memorial Sloan Kettering Cancer Center, New York, NY (S.C.).
¹⁴ Department of Ophthalmology, Eugene and Marilyn Glick Eye Institute, Indiana University School of Medicine, Indianapolis. (S.L.C., M.B.G.).
¹⁵ Center for Immunobiology, Department of Investigative Medicine, Western Michigan University Homer Stryker MD School of Medicine, Kalamazoo (M. Yoshimoto). Department of Ophthalmology, University of Alabama at Birmingham (S.L.C., M.B.G.).
¹⁶ Department of Anatomy, Cell Biology & Physiology, Indiana University School of Medicine, Indianapolis. (D.P.B.).
¹⁷ Department of Surgery, McGowan Institute for Regenerative Medicine, University of Pittsburgh, PA (M.C.Y.).

^# Contributed equally.

PMID: 38682338
DOI: 10.1161/CIRCULATIONAHA.122.061833

Abstract

Background: Most organs are maintained lifelong by resident stem/progenitor cells. During development and regeneration, lineage-specific stem/progenitor cells can contribute to the growth or maintenance of different organs, whereas fully differentiated mature cells have less regenerative potential. However, it is unclear whether vascular endothelial cells (ECs) are also replenished by stem/progenitor cells with EC-repopulating potential residing in blood vessels. It has been reported recently that some EC populations possess higher clonal proliferative potential and vessel-forming capacity compared with mature ECs. Nevertheless, a marker to identify vascular clonal repopulating ECs (CRECs) in murine and human individuals is lacking, and, hence, the mechanism for the proliferative, self-renewal, and vessel-forming potential of CRECs is elusive.

Methods: We analyzed colony-forming, self-renewal, and vessel-forming potential of ABCG2 (ATP binding cassette subfamily G member 2)-expressing ECs in human umbilical vessels. To study the contribution of Abcg2-expressing ECs to vessel development and regeneration, we developed Abcg2Cre^Ert2;ROSA TdTomato mice and performed lineage tracing during mouse development and during tissue regeneration after myocardial infarction injury. RNA sequencing and chromatin methylation chromatin immunoprecipitation followed by sequencing were conducted to study the gene regulation in Abcg2-expressing ECs.

Results: In human and mouse vessels, ECs with higher ABCG2 expression (ABCECs) possess higher clonal proliferative potential and in vivo vessel-forming potential compared with mature ECs. These cells could clonally contribute to vessel formation in primary and secondary recipients after transplantation. These features of ABCECs meet the criteria of CRECs. Results from lineage tracing experiments confirm that Abcg2-expressing CRECs (AbcCRECs) contribute to arteries, veins, and capillaries in cardiac tissue development and vascular tissue regeneration after myocardial infarction. Transcriptome and epigenetic analyses reveal that a gene expression signature involved in angiogenesis and vessel development is enriched in AbcCRECs. In addition, various angiogenic genes, such as Notch2 and Hey2, are bivalently modified by trimethylation at the 4th and 27th lysine residue of histone H3 (H3K4me3 and H3K27me3) in AbcCRECs.

Conclusions: These results are the first to establish that a single prospective marker identifies CRECs in mice and human individuals, which holds promise to provide new cell therapies for repair of damaged vessels in patients with endothelial dysfunction.

Keywords: angiogenesis; blood vessels; developmental biology; endothelial cells.

Abstract

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