Localized enteric neural deficiency and reduced nerve-mediated anorectal contractility in immunodeficient RetS812F/+ mice

Evaluating cell-based therapies for the enteric nervous system (ENS) requires an in vivo environment with a localized enteric neural-deficient region. However, preclinical studies using human cells are limited by the lack of suitable animal models; conventional aganglionic models often suffer from early lethality, and there are few immunodeficient strains with a defined enteric neural-deficient segment for xenotransplantation. In this study, we established the B6RGS-RetS812F/+ mouse, a novel immunodeficient strain with a localized deficiency of enteric nerves and reduced nerve-related anorectal contraction.

While these mice did not exhibit overt defecation defects or stool congestion, this phenotype allowed for the survival period necessary to perform transplantation experiments. Using the B6RGS background, we observed the short-term retention of human induced pluripotent stem cell-derived vagal neural crest cells within the distal anorectum. Our results suggest that the B6RGS-RetS812F/+ mouse may serve as a practical recipient for evaluating the initial engraftment of human-derived cell therapies for enteric nerve deficiencies.

B6.Cg-Sirpa Rag2 Il2rg/Jic (B6RGS) mice in this study are available from the Central Institute for Experimental Medicine and Life Science (CIEM). To generate B6RGS-RetS812F/+ mice, frozen sperm from male C57BL/6N-RetS812F/+ mice should be obtained from Kobe University. The parent 1231A3 human-induced pluripotent stem cell lines should be provided from Yamanaka laboratory.

Requests for further resources should be directed to and will be fulfilled by the lead contact, Makoto Ikeya ([email protected]). We sincerely thank the former and current members of the T-CiRA Ikeya project, especially Yayoi Toyooka, Daisuke Kamiya, Teppei Akaboshi, and Hideyuki Hiyoshi, for their valuable support; Mika Sato and Yoshiki Yamaguchi from Orizuru Therapeutics for their technical assistance; and Yoshiaki Kassai, Yasushi Kajii, and Shinya Yamanaka for critically evaluating this study. T-CiRA Discovery and Innovation, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan Yusaku Komoike, Teruyoshi Yamashita, Taiki Nakajima & Hirokazu Matsumoto Takeda-CiRA Joint Program (T-CiRA), Fujisawa, Kanagawa, Japan Yusaku Komoike, Teruyoshi Yamashita, Taiki Nakajima, Hirokazu Matsumoto & Makoto Ikeya Central Institute for Experimental Medicine and Life Science (CIEM), Kawasaki, Kanagawa, Japan Division of Neural Differentiation and Regeneration, Department of Physiology and Cell Biology, Kobe University Graduate School of Medicine, Kobe, Hyogo, Japan Department of Clinical Application, Center for iPS Cell Research and Application (CiRA), Kyoto University, Sakyo-ku, Kyoto, 606-8507, Japan Yusaku Komoike, Teruyoshi Yamashita, Taiki Nakajima and Hirokazu Matsumoto are employees of Takeda Pharmaceutical Company Limited.

Motohito Goto and Masafumi Yamamoto are employees of Central Institute for Experimental Medicine and Life Science. Financial support was provided by Takeda Pharmaceutical Co Ltd. The other authors declare no competing interests.

This study was supported by a T-CiRA budget from Takeda Pharmaceutical Company, Limited. All authors had access to the study data and reviewed and approved the final manuscript. Requests for further information should be directed to the lead contact, Makoto Ikeya ([email protected]).