IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B

Louis R Parham; Patrick A Williams; Kay Katada; Shaneice K Nettleford; Priya Chatterji; Kofi K Acheampong; Charles H Danan; Xianghui Ma; Lauren A Simon; Kaitlyn E Naughton; Rei Mizuno; Tatiana Karakasheva; Emily A McMillan; Kelly A Whelan; Donita C Brady; Sydney M Shaffer; Kathryn E Hamilton

doi:10.1016/j.jcmgh.2023.12.001

IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B

Louis R Parham, Patrick A Williams, Kay Katada, Shaneice K Nettleford, Priya Chatterji, Kofi K Acheampong, Charles H Danan, Xianghui Ma, Lauren A Simon, Kaitlyn E Naughton, Rei Mizuno, Tatiana Karakasheva, Emily A McMillan, Kelly A Whelan, Donita C Brady, Sydney M Shaffer, Kathryn E Hamilton

Cancer Signaling and Microenvironment (CSM)

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

BACKGROUND & AIMS: The intestinal epithelium interfaces with a diverse milieu of luminal contents while maintaining robust digestive and barrier functions. Facultative intestinal stem cells are cells that survive tissue injury and divide to re-establish the epithelium. Prior studies have shown autophagic state as functional marker of facultative intestinal stem cells, but regulatory mechanisms are not known. The current study evaluated a post-transcriptional regulation of autophagy as an important factor for facultative stem cell state and tissue regeneration.

METHODS: We evaluated stem cell composition, autophagic vesicle content, organoid formation, and in vivo regeneration in mice with intestinal epithelial deletion of the RNA binding protein IGF2 messenger RNA binding protein 1 (IMP1). The contribution of autophagy to resulting in vitro and in vivo phenotypes was evaluated via genetic inactivation of Atg7. Molecular analyses of IMP1 modulation of autophagy at the protein and transcript localization levels were performed using IMP1 mutant studies and single-molecule fluorescent in situ hybridization.

RESULTS: Epithelial Imp1 deletion reduced leucine rich repeat containing G protein coupled receptor 5 cell frequency but enhanced both organoid formation efficiency and in vivo regeneration after irradiation. We confirmed prior studies showing increased autophagy with IMP1 deletion. Deletion of Atg7 reversed the enhanced regeneration observed with Imp1 deletion. IMP1 deletion or mutation of IMP1 phosphorylation sites enhanced expression of essential autophagy protein microtubule-associated protein 1 light chain 3β. Furthermore, immunofluorescence imaging coupled with single-molecule fluorescent in situ hybridization showed IMP1 colocalization with MAP1LC3B transcripts at homeostasis. Stress induction led to decreased colocalization.

CONCLUSIONS: Depletion of IMP1 enhances autophagy, which promotes intestinal regeneration via expansion of facultative intestinal stem cells.

Original language	English
Pages (from-to)	439-451
Number of pages	13
Journal	Cellular and Molecular Gastroenterology and Hepatology
Volume	17
Issue number	3
Early online date	Nov 9 2023
DOIs	https://doi.org/10.1016/j.jcmgh.2023.12.001
State	Published - 2024

Keywords

regeneration
post-transcriptional gene regulation
intestinal homeostasis
Autophagy
Intestines
Animals
Intestinal Mucosa/metabolism
In Situ Hybridization, Fluorescence
Mice
RNA-Binding Proteins/genetics
Stem Cells/metabolism

Access to Document

10.1016/j.jcmgh.2023.12.001

Cite this

Parham, L. R., Williams, P. A., Katada, K., Nettleford, S. K., Chatterji, P., Acheampong, K. K., Danan, C. H., Ma, X., Simon, L. A., Naughton, K. E., Mizuno, R., Karakasheva, T., McMillan, E. A., Whelan, K. A., Brady, D. C., Shaffer, S. M., & Hamilton, K. E. (2024). IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B. Cellular and Molecular Gastroenterology and Hepatology, 17(3), 439-451. https://doi.org/10.1016/j.jcmgh.2023.12.001

@article{019429d4a2884a1ab40ea6b41fdf0962,

title = "IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B",

abstract = "BACKGROUND & AIMS: The intestinal epithelium interfaces with a diverse milieu of luminal contents while maintaining robust digestive and barrier functions. Facultative intestinal stem cells are cells that survive tissue injury and divide to re-establish the epithelium. Prior studies have shown autophagic state as functional marker of facultative intestinal stem cells, but regulatory mechanisms are not known. The current study evaluated a post-transcriptional regulation of autophagy as an important factor for facultative stem cell state and tissue regeneration.METHODS: We evaluated stem cell composition, autophagic vesicle content, organoid formation, and in vivo regeneration in mice with intestinal epithelial deletion of the RNA binding protein IGF2 messenger RNA binding protein 1 (IMP1). The contribution of autophagy to resulting in vitro and in vivo phenotypes was evaluated via genetic inactivation of Atg7. Molecular analyses of IMP1 modulation of autophagy at the protein and transcript localization levels were performed using IMP1 mutant studies and single-molecule fluorescent in situ hybridization.RESULTS: Epithelial Imp1 deletion reduced leucine rich repeat containing G protein coupled receptor 5 cell frequency but enhanced both organoid formation efficiency and in vivo regeneration after irradiation. We confirmed prior studies showing increased autophagy with IMP1 deletion. Deletion of Atg7 reversed the enhanced regeneration observed with Imp1 deletion. IMP1 deletion or mutation of IMP1 phosphorylation sites enhanced expression of essential autophagy protein microtubule-associated protein 1 light chain 3β. Furthermore, immunofluorescence imaging coupled with single-molecule fluorescent in situ hybridization showed IMP1 colocalization with MAP1LC3B transcripts at homeostasis. Stress induction led to decreased colocalization.CONCLUSIONS: Depletion of IMP1 enhances autophagy, which promotes intestinal regeneration via expansion of facultative intestinal stem cells.",

keywords = "regeneration, post-transcriptional gene regulation, intestinal homeostasis, Autophagy, Intestines, Animals, Intestinal Mucosa/metabolism, In Situ Hybridization, Fluorescence, Mice, RNA-Binding Proteins/genetics, Stem Cells/metabolism",

author = "Parham, {Louis R} and Williams, {Patrick A} and Kay Katada and Nettleford, {Shaneice K} and Priya Chatterji and Acheampong, {Kofi K} and Danan, {Charles H} and Xianghui Ma and Simon, {Lauren A} and Naughton, {Kaitlyn E} and Rei Mizuno and Tatiana Karakasheva and McMillan, {Emily A} and Whelan, {Kelly A} and Brady, {Donita C} and Shaffer, {Sydney M} and Hamilton, {Kathryn E}",

year = "2024",

doi = "10.1016/j.jcmgh.2023.12.001",

language = "English",

volume = "17",

pages = "439--451",

journal = "Cellular and Molecular Gastroenterology and Hepatology",

issn = "2352-345X",

publisher = "Elsevier Inc.",

number = "3",

}

Parham, LR, Williams, PA, Katada, K, Nettleford, SK, Chatterji, P, Acheampong, KK, Danan, CH, Ma, X, Simon, LA, Naughton, KE, Mizuno, R, Karakasheva, T, McMillan, EA, Whelan, KA, Brady, DC, Shaffer, SM & Hamilton, KE 2024, 'IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B', Cellular and Molecular Gastroenterology and Hepatology, vol. 17, no. 3, pp. 439-451. https://doi.org/10.1016/j.jcmgh.2023.12.001

TY - JOUR

T1 - IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B

AU - Parham, Louis R

AU - Williams, Patrick A

AU - Katada, Kay

AU - Nettleford, Shaneice K

AU - Chatterji, Priya

AU - Acheampong, Kofi K

AU - Danan, Charles H

AU - Ma, Xianghui

AU - Simon, Lauren A

AU - Naughton, Kaitlyn E

AU - Mizuno, Rei

AU - Karakasheva, Tatiana

AU - McMillan, Emily A

AU - Whelan, Kelly A

AU - Brady, Donita C

AU - Shaffer, Sydney M

AU - Hamilton, Kathryn E

PY - 2024

Y1 - 2024

N2 - BACKGROUND & AIMS: The intestinal epithelium interfaces with a diverse milieu of luminal contents while maintaining robust digestive and barrier functions. Facultative intestinal stem cells are cells that survive tissue injury and divide to re-establish the epithelium. Prior studies have shown autophagic state as functional marker of facultative intestinal stem cells, but regulatory mechanisms are not known. The current study evaluated a post-transcriptional regulation of autophagy as an important factor for facultative stem cell state and tissue regeneration.METHODS: We evaluated stem cell composition, autophagic vesicle content, organoid formation, and in vivo regeneration in mice with intestinal epithelial deletion of the RNA binding protein IGF2 messenger RNA binding protein 1 (IMP1). The contribution of autophagy to resulting in vitro and in vivo phenotypes was evaluated via genetic inactivation of Atg7. Molecular analyses of IMP1 modulation of autophagy at the protein and transcript localization levels were performed using IMP1 mutant studies and single-molecule fluorescent in situ hybridization.RESULTS: Epithelial Imp1 deletion reduced leucine rich repeat containing G protein coupled receptor 5 cell frequency but enhanced both organoid formation efficiency and in vivo regeneration after irradiation. We confirmed prior studies showing increased autophagy with IMP1 deletion. Deletion of Atg7 reversed the enhanced regeneration observed with Imp1 deletion. IMP1 deletion or mutation of IMP1 phosphorylation sites enhanced expression of essential autophagy protein microtubule-associated protein 1 light chain 3β. Furthermore, immunofluorescence imaging coupled with single-molecule fluorescent in situ hybridization showed IMP1 colocalization with MAP1LC3B transcripts at homeostasis. Stress induction led to decreased colocalization.CONCLUSIONS: Depletion of IMP1 enhances autophagy, which promotes intestinal regeneration via expansion of facultative intestinal stem cells.

AB - BACKGROUND & AIMS: The intestinal epithelium interfaces with a diverse milieu of luminal contents while maintaining robust digestive and barrier functions. Facultative intestinal stem cells are cells that survive tissue injury and divide to re-establish the epithelium. Prior studies have shown autophagic state as functional marker of facultative intestinal stem cells, but regulatory mechanisms are not known. The current study evaluated a post-transcriptional regulation of autophagy as an important factor for facultative stem cell state and tissue regeneration.METHODS: We evaluated stem cell composition, autophagic vesicle content, organoid formation, and in vivo regeneration in mice with intestinal epithelial deletion of the RNA binding protein IGF2 messenger RNA binding protein 1 (IMP1). The contribution of autophagy to resulting in vitro and in vivo phenotypes was evaluated via genetic inactivation of Atg7. Molecular analyses of IMP1 modulation of autophagy at the protein and transcript localization levels were performed using IMP1 mutant studies and single-molecule fluorescent in situ hybridization.RESULTS: Epithelial Imp1 deletion reduced leucine rich repeat containing G protein coupled receptor 5 cell frequency but enhanced both organoid formation efficiency and in vivo regeneration after irradiation. We confirmed prior studies showing increased autophagy with IMP1 deletion. Deletion of Atg7 reversed the enhanced regeneration observed with Imp1 deletion. IMP1 deletion or mutation of IMP1 phosphorylation sites enhanced expression of essential autophagy protein microtubule-associated protein 1 light chain 3β. Furthermore, immunofluorescence imaging coupled with single-molecule fluorescent in situ hybridization showed IMP1 colocalization with MAP1LC3B transcripts at homeostasis. Stress induction led to decreased colocalization.CONCLUSIONS: Depletion of IMP1 enhances autophagy, which promotes intestinal regeneration via expansion of facultative intestinal stem cells.

KW - regeneration

KW - post-transcriptional gene regulation

KW - intestinal homeostasis

KW - Autophagy

KW - Intestines

KW - Animals

KW - Intestinal Mucosa/metabolism

KW - In Situ Hybridization, Fluorescence

KW - Mice

KW - RNA-Binding Proteins/genetics

KW - Stem Cells/metabolism

UR - https://pubmed.ncbi.nlm.nih.gov/38081361/

UR - http://www.scopus.com/inward/record.url?scp=85183173731&partnerID=8YFLogxK

U2 - 10.1016/j.jcmgh.2023.12.001

DO - 10.1016/j.jcmgh.2023.12.001

M3 - Article

C2 - 38081361

SN - 2352-345X

VL - 17

SP - 439

EP - 451

JO - Cellular and Molecular Gastroenterology and Hepatology

JF - Cellular and Molecular Gastroenterology and Hepatology

IS - 3

ER -

IGF2BP1/IMP1 Deletion Enhances a Facultative Stem Cell State via Regulation of MAP1LC3B

Abstract

Keywords

Access to Document

Other files and links

Fingerprint

Cite this