Original Research
Insulin-like Growth Factor-1 and mTORC1 Signaling Promote the Intestinal Regenerative Response After Irradiation Injury

https://doi.org/10.1016/j.jcmgh.2020.05.013Get rights and content
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Background & Aims

Intestinal crypts have a remarkable capacity to regenerate after injury from loss of crypt base columnar (CBC) stem cells. After injury, facultative stem cells (FSCs) are activated to replenish the epithelium and replace lost CBCs. Our aim was to assess the role of insulin-like growth factor-1 (IGF-1) to activate FSCs for crypt repair.

Methods

The intestinal regenerative response was measured after whole body 12-Gy γ-irradiation of adult mice. IGF-1 signaling or its downstream effector mammalian target of rapamycin complex 1 (mTORC1) was inhibited by administering BMS-754807 or rapamycin, respectively. Mice with inducible Rptor gene deletion were studied to test the role of mTORC1 signaling in the intestinal epithelium. FSC activation post-irradiation was measured by lineage tracing.

Results

We observed a coordinate increase in growth factor expression, including IGF-1, at 2 days post-irradiation, followed by a surge in mTORC1 activity during the regenerative phase of crypt repair at day 4. IGF-1 was localized to pericryptal mesenchymal cells, and IGF-1 receptor was broadly expressed in crypt progenitor cells. Inhibition of IGF-1 signaling via BMS-754807 treatment impaired crypt regeneration after 12-Gy irradiation, with no effect on homeostasis. Similarly, rapamycin inhibition of mTORC1 during the growth factor surge blunted the regenerative response. Analysis of Villin-CreERT2;Rptorfl/fl mice showed that epithelial mTORC1 signaling was essential for crypt regeneration. Lineage tracing from Bmi1-marked cells showed that rapamycin blocked FSC activation post-irradiation.

Conclusions

Our study shows that IGF-1 signaling through mTORC1 drives crypt regeneration. We propose that IGF-1 release from pericryptal cells stimulates mTORC1 in FSCs to regenerate lost CBCs.

Keywords

IGF-1
Raptor
Rapamycin
Intestinal Repair
Crypt Regeneration
Intestinal Stem Cells

Abbreviations used in this paper

ANOVA
analysis of variance
CBC
crypt base columnar
DPI
days post-irradiation
EDU
5-ethynyl-2´-deoxyuridine
FSC
facultative stem cell
HPI
hours post-irradiation
IGF-1
insulin-like growth factor-1
ISC
intestinal stem cell
mTORC1
mammalian target of rapamycin complex 1
PBS
phosphate-buffered saline
qPCR
quantitative reverse transcriptase polymerase chain reaction
SEM
standard error of the mean
UNIRR
unirradiated controls

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Conflicts of interest The authors disclose no conflicts.

Funding NB was supported by the Cellular & Molecular Biology program, a Rackham International Student Fellowship, and the Bernard L. Maas Fellowship; KPM was supported by the Training Program in Organogenesis NIH T32 HD007505. The research was funded by NIH R01-DK118023 to LCS, R03-DK114656 to KSY, BWF CAMS to KSY, and Core support from the Michigan Gastrointestinal Research Center Grant NIH P30-DK34933.