Wnt-induced, TRP53-mediated Cell Cycle Arrest of Precursors Underlies Interstitial Cell of Cajal Depletion During Aging

Yujiro Hayashi; David T Asuzu; Michael R Bardsley; Gabriella B Gajdos; Sergiy M Kvasha; David R Linden; Rea A Nagy; Siva Arumugam Saravanaperumal; Sabriya A Syed; Yoshitaka Toyomasu; Huihuang Yan; Eduardo N Chini; Simon J Gibbons; Todd A Kellogg; Khashayarsha Khazaie; Makoto Kuro-O; Jair Machado Espindola Netto; Mahendra Pal Singh; James G Tidball; Michelle Wehling-Henricks; Gianrico Farrugia; Tamas Ordog

doi:10.1016/j.jcmgh.2020.07.011

Wnt-induced, TRP53-mediated Cell Cycle Arrest of Precursors Underlies Interstitial Cell of Cajal Depletion During Aging

Cell Mol Gastroenterol Hepatol. 2021;11(1):117-145. doi: 10.1016/j.jcmgh.2020.07.011. Epub 2020 Aug 7.

Authors

Yujiro Hayashi¹, David T Asuzu², Michael R Bardsley², Gabriella B Gajdos², Sergiy M Kvasha², David R Linden³, Rea A Nagy², Siva Arumugam Saravanaperumal², Sabriya A Syed², Yoshitaka Toyomasu², Huihuang Yan⁴, Eduardo N Chini⁵, Simon J Gibbons³, Todd A Kellogg⁶, Khashayarsha Khazaie⁷, Makoto Kuro-O⁸, Jair Machado Espindola Netto⁵, Mahendra Pal Singh⁷, James G Tidball⁹, Michelle Wehling-Henricks⁹, Gianrico Farrugia³, Tamas Ordog¹⁰

Affiliations

¹ Enteric Neuroscience Program and Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota; Gastroenterology Research Unit, Division of Gastroenterology and Hepatology, Department of Medicine, Mayo Clinic, Rochester, Minnesota. Electronic address: hayashi.yujiro@mayo.edu.
² Enteric Neuroscience Program and Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota; Gastroenterology Research Unit, Division of Gastroenterology and Hepatology, Department of Medicine, Mayo Clinic, Rochester, Minnesota.
³ Enteric Neuroscience Program and Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota.
⁴ Division of Biomedical Statistics and Informatics, Department of Health Sciences Research, Mayo Clinic, Rochester, Minnesota.
⁵ Signal Transduction and Molecular Nutrition Laboratory, Kogod Aging Center and Department of Anesthesiology and Perioperative Medicine, Mayo Clinic, Rochester, Minnesota.
⁶ Department of Surgery, Mayo Clinic, Rochester, Minnesota.
⁷ Department of Immunology, Mayo Clinic, Rochester, Minnesota.
⁸ Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas; Division of Anti-aging Medicine, Center for Molecular Medicine, Jichi Medical University, Shimotsuke, Tochigi, Japan.
⁹ Department of Integrative Biology and Physiology, University of California, Los Angeles, California.
¹⁰ Enteric Neuroscience Program and Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota; Gastroenterology Research Unit, Division of Gastroenterology and Hepatology, Department of Medicine, Mayo Clinic, Rochester, Minnesota; Center for Individualized Medicine, Mayo Clinic, Rochester, Minnesota. Electronic address: ordog.tamas@mayo.edu.

Abstract

Background & aims: Gastric dysfunction in the elderly may cause reduced food intake, frailty, and increased mortality. The pacemaker and neuromodulator cells interstitial cells of Cajal (ICC) decline with age in humans, and their loss contributes to gastric dysfunction in progeric klotho mice hypomorphic for the anti-aging Klotho protein. The mechanisms of ICC depletion remain unclear. Klotho attenuates Wnt (wingless-type MMTV integration site) signaling. Here, we examined whether unopposed Wnt signaling could underlie aging-associated ICC loss by up-regulating transformation related protein TRP53 in ICC stem cells (ICC-SC).

Methods: Mice aged 1-107 weeks, klotho mice, APC^Δ468 mice with overactive Wnt signaling, mouse ICC-SC, and human gastric smooth muscles were studied by RNA sequencing, reverse transcription-polymerase chain reaction, immunoblots, immunofluorescence, histochemistry, flow cytometry, and methyltetrazolium, ethynyl/bromodeoxyuridine incorporation, and ex-vivo gastric compliance assays. Cells were manipulated pharmacologically and by gene overexpression and RNA interference.

Results: The klotho and aged mice showed similar ICC loss and impaired gastric compliance. ICC-SC decline preceded ICC depletion. Canonical Wnt signaling and TRP53 increased in gastric muscles of klotho and aged mice and middle-aged humans. Overstimulated canonical Wnt signaling increased DNA damage response and TRP53 and reduced ICC-SC self-renewal and gastric ICC. TRP53 induction persistently inhibited G₁/S and G₂/M cell cycle phase transitions without activating apoptosis, autophagy, cellular quiescence, or canonical markers/mediators of senescence. G₁/S block reflected increased cyclin-dependent kinase inhibitor 1B and reduced cyclin D1 from reduced extracellular signal-regulated kinase activity.

Conclusions: Increased Wnt signaling causes age-related ICC loss by up-regulating TRP53, which induces persistent ICC-SC cell cycle arrest without up-regulating canonical senescence markers.

Keywords: Compliance; Senescence; Stem Cell.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Adenomatous Polyposis Coli Protein / genetics
Aging / physiology*
Animals
Cell Cycle Checkpoints
Cellular Senescence / physiology*
Female
Humans
Interstitial Cells of Cajal / physiology*
Klotho Proteins / genetics
Male
Mice
Mice, Transgenic
Middle Aged
Models, Animal
Stomach / cytology
Stomach / physiology*
Tumor Suppressor Protein p53 / metabolism
Up-Regulation
Wnt Signaling Pathway
Young Adult

Substances

Adenomatous Polyposis Coli Protein
TP53 protein, human
Trp53 protein, mouse
Tumor Suppressor Protein p53
adenomatous polyposis coli protein, mouse
Klotho Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding