bims-instec 2021-09-26 papers

bims-instec

Biomed News

on Intestinal stem cells and chemoresistance in colon cancer and intestinal regeneration

Issue of 2021–09–26
eight papers selected by
Maria-Virginia Giolito, IRFAC/UMR-S1113 INSERM

Neurotensin regulates proliferation and stem cell function in the small intestine in a nutrient-dependent manner.
A High-Fat Diet Controls MHC Class II on ISCs via the Microbiome.
Pharmacological targeting PTK6 inhibits the JAK2/STAT3 sustained stemness and reverses chemoresistance of colorectal cancer.
Germline mutations in a DNA repair pathway are associated with familial colorectal cancer.
SMAR1 Suppresses the Cancer Stem Cell Population via hTERT Repression in Colorectal Cancer Cells.
Multiscale analysis reveals that diet-dependent midgut plasticity emerges from alterations in both stem cell niche coupling and enterocyte size.
HOPX+ injury-resistant intestinal stem cells drive epithelial recovery after severe intestinal ischemia.
Diet-induced alteration of intestinal stem cell function underlies obesity and prediabetes in mice.

Cell Mol Gastroenterol Hepatol. 2021 Sep 21. pii: S2352-345X(21)00194-6. [Epub ahead of print]

Neurotensin regulates proliferation and stem cell function in the small intestine in a nutrient-dependent manner.

Stephanie A Rock, Kai Jiang, Yuanyuan Wu, Yajuan Liu, Jing Li, Heidi L Weiss, Chi Wang, Jianhang Jia, Tianyan Gao, B Mark Evers.

   BACKGROUND & AIMS: Intestinal stem cells (ISCs) are sensitive to dietary alterations and nutrient availability. Neurotensin (NT), a gut peptide localized predominantly to the small bowel and released by fat ingestion, stimulates the growth of intestinal mucosa under basal conditions and during periods of nutrient deprivation, suggesting a possible role for NT on ISC function.
METHODS: Lgr5-EGFP, NT wild type (Nt+/+) and Lgr5-EGFP, NT knockout (Nt-/-) mice were fed ad libitum (AL) or fasted for 48 h. Small intestine tissue and crypts were examined by gene expression analyses, fluorescence-activated cell sorting, western blot, immunohistochemistry, and crypt-derived organoid culture. Drosophila expressing NT in midgut enteroendocrine cells were fed a standard diet or low-energy diet and esg-GFP+ ISCs quantified via immunofluorescence.
RESULTS: Loss of NT impaired crypt cell proliferation and ISC function in a manner dependent on nutrient status. Under nutrient-rich conditions, NT stimulated ERK1/2 signaling and the expression of genes that promote cell cycle progression, leading to crypt cell proliferation. Under conditions of nutrient depletion, NT stimulated WNT/®-catenin signaling and promoted an ISC gene signature, leading to enhanced ISC function. NT was required for the induction of WNT/®-catenin signaling and ISC-specific gene expression during nutrient depletion, and loss of NT reduced crypt cell proliferation and impaired ISC function and Lgr5 expression in the intestine during fasting. Conversely, the expression of NT in midgut enteroendocrine cells of Drosophila prevented loss of ISCs during nutrient depletion.
CONCLUSION: Collectively, our findings establish an evolutionarily conserved role for NT in ISC maintenance during nutritional stress.

Keywords:  Drosophila; Neurotensin; diet; intestinal stem cells

DOI:  https://doi.org/10.1016/j.jcmgh.2021.09.006
Cancer Discov. 2021 Sep 24.

A High-Fat Diet Controls MHC Class II on ISCs via the Microbiome.

A high-fat diet (HFD) increases tumorigenicity by reducing MHC class II on intestinal stem cells (ISC).

DOI: https://doi.org/10.1158/2159-8290.CD-RW2021-134
J Exp Clin Cancer Res. 2021 Sep 22. 40(1): 297

Pharmacological targeting PTK6 inhibits the JAK2/STAT3 sustained stemness and reverses chemoresistance of colorectal cancer.

Chaoqun Liu, Zhihua Pan, Qian Chen, Zetao Chen, Weiwei Liu, Ling Wu, Muhong Jiang, Wandie Lin, Yujie Zhang, Weihao Lin, Rui Zhou, Liang Zhao.

   BACKGROUND: Chemoresistance is the major cause of chemotherapy failure in patients with colorectal cancer (CRC). Protein tyrosine kinase 6 (PTK6) is aberrantly overexpressed in clinical CRC tissues undergoing chemotherapy. We studied if PTK6 contributed to the chemoresistance of CRC in human and mice.
METHODS: We obtained tissue samples from patients with CRC and measured the expression of PTK6 by immunohistochemistry. Gain- and loss-of-function assays were performed to study the biological functions of PTK6. We constructed the FLAG-tagged wild type (WT), kinase-dead, and inhibition-defective recombinant mutants of PTK6 to study the effect phosphorylated activation of PTK6 played on CRC cell stemness and chemoresistance. We used small molecule inhibitor XMU-MP-2 to test the influence of PTK6 on sensitivity of CRC cells to 5-FU/L-OHP in both nude mouse and patient-derived xenograft (PDX) animal models.
RESULTS: PTK6 is overexpressed in CRC tissues and plays a stimulatory role in the proliferation and chemoresistance of CRC cells both in vitro and in vivo. PTK6, especially the phosphorylated PTK6, can promote the stemness of CRC cells through interacting with JAK2 and phosphorylating it to activate the JAK2/STAT3 signaling. Pharmacological inhibition of PTK6 using XMU-MP-2 effectively reduces the stemness property of CRC cells and improves its chemosensitivity to 5-FU/L-OHP in both nude mice subcutaneously implanted tumor model and PDX model constructed with NOD-SCID mice.
CONCLUSIONS: PTK6 interacts with JAK2 and phosphorylates it to activate JAK2/STAT3 signaling to promote the stemness and chemoresistance of CRC cells. Pharmacological inhibition of PTK6 by small molecule inhibitor dramatically enhances the sensitivity to chemotherapy in nude mice and PDX models.

Keywords:  Chemoresistance; Colorectal cancer; PTK6; Small molecule kinase inhibitor; Stemness

DOI:  https://doi.org/10.1186/s13046-021-02059-6
JCI Insight. 2021 Sep 22. pii: e148931. [Epub ahead of print]6(18):

Germline mutations in a DNA repair pathway are associated with familial colorectal cancer.

Pingping Xu, Danfeng Sun, Yaqi Gao, Yi Jiang, Ming Zhong, Gang Zhao, Jinxian Chen, Zheng Wang, Qiang Liu, Jie Hong, Haoyan Chen, Ying-Xuan Chen, Jing-Yuan Fang.

  Aiming to identify rare high-penetrance mutations in new genes for the underlying predisposition in familial colorectal cancer (CRC), we performed whole-exome sequencing in 24 familial CRCs. Mutations in genes that regulate DNA repair (RMI1, PALB2, FANCI) were identified that were related to the Fanconi anemia DNA repair pathway. In one pedigree, we found a nonsense mutation in CHEK2. CHEK2 played an essential role in cell cycle and DNA damage repair. Somatic mutation analysis in CHEK2 variant carriers showed mutations in TP53, APC, and FBXW7. Loss of heterozygosity was found in carcinoma of CHEK2 variant carrier, and IHC showed loss of Chk2 expression in cancer tissue. We identified a second variant in CHEK2 in 126 sporadic CRCs. A KO cellular model for CHEK2 (CHEK2KO) was generated by CRISPR/Cas9. Functional experiments demonstrated that CHEK2KO cells showed defective cell cycle arrest and apoptosis, as well as reduced p53 phosphorylation, upon DNA damage. We associated germline mutations in genes that regulate the DNA repair pathway with the development of CRC. We identified CHEK2 as a regulator of DNA damage response and perhaps as a gene involved in CRC germline predisposition. These findings link CRC predisposition to the DNA repair pathway, supporting the connection between genome integrity and cancer risk.

Keywords:  Cancer; Cell cycle; DNA repair; Gastroenterology; Genetics

DOI:  https://doi.org/10.1172/jci.insight.148931
Int J Biochem Cell Biol. 2021 Sep 19. pii: S1357-2725(21)00166-7. [Epub ahead of print] 106085

SMAR1 Suppresses the Cancer Stem Cell Population via hTERT Repression in Colorectal Cancer Cells.

Apoorva Parulekar, Arpankumar Choksi, Nandaraj Taye, Kumar V S Totakura, Priyanka Firmal, Gopal C Kundu, Samit Chattopadhyay.

  One of the hallmarks of a cancer cell is the ability for indefinite proliferation leading to the immortalization of the cell. Activation of several signaling pathways leads to the immortalization of cancer cells via the reactivation of enzyme telomerase (hTERT). hTERT is active in germ cells, stem cells and also cancer cells. An earlier report from our lab suggests that SMAR1, a tumor suppressor protein, is significantly downregulated in the higher grades of colorectal cancers. Our study identifies SMAR1 as a transcriptional repressor of hTERT. We find that SMAR1 interacts with HDAC1/mSin3a co-repressor complex at the hTERT promoter and brings about HDAC1-mediated transcriptional repression of the promoter. Most solid tumors including colorectal cancer reactivate hTERT expression as it confers several advantages to the cancer cells like increased proliferation and angiogenesis. One of these non-canonical functions of hTERT is inducing the pool of cancer stem cell population. We find that in the CD133HighCD44High cancer stem cells population, SMAR1 expression is highly diminished leading to elevated hTERT expression. We also find that knockdown of SMAR1 promotes total CD133+CD44+ population and impart enhanced sphere-forming ability to the colorectal cancer cells. SMAR1 also inhibits invasion and metastasis in colorectal cancer cell lines via repression of hTERT. Our study provides evidence that downregulation of SMAR1 causes activation of hTERT leading to an increase in the cancer stem cell phenotype in colorectal cancer cells.

Keywords:  CD133; CD44; CSC; HDAC1; SMAR1; hTERT promoter

DOI:  https://doi.org/10.1016/j.biocel.2021.106085
Elife. 2021 Sep 23. pii: e64125. [Epub ahead of print]10

Multiscale analysis reveals that diet-dependent midgut plasticity emerges from alterations in both stem cell niche coupling and enterocyte size.

Alessandro Bonfini, Adam J Dobson, David Duneau, Jonathan Revah, Xi Liu, Philip Houtz, Nicolas Buchon.

  The gut is the primary interface between an animal and food, but how it adapts to qualitative dietary variation is poorly defined. We find that the Drosophila midgut plastically resizes following changes in dietary composition. A panel of nutrients collectively promote gut growth, which sugar opposes. Diet influences absolute and relative levels of enterocyte loss and stem cell proliferation, which together determine cell numbers. Diet also influences enterocyte size. A high sugar diet inhibits translation and uncouples ISC proliferation from expression of niche-derived signals but, surprisingly, rescuing these effects genetically was not sufficient to modify diet's impact on midgut size. However, when stem cell proliferation was deficient, diet's impact on enterocyte size was enhanced, and reducing enterocyte-autonomous TOR signaling was sufficient to attenuate diet-dependent midgut resizing. These data clarify the complex relationships between nutrition, epithelial dynamics, and cell size, and reveal a new mode of plastic, diet-dependent organ resizing.

Keywords:  D. melanogaster; developmental biology; regenerative medicine; stem cells

DOI:  https://doi.org/10.7554/eLife.64125
Am J Physiol Gastrointest Liver Physiol. 2021 Sep 22.

HOPX+ injury-resistant intestinal stem cells drive epithelial recovery after severe intestinal ischemia.

Amy Stieler Stewart, Cecilia Renee Schaaf, Jennifer A Luff, John M Freund, Thomas C Becker, Sara R Tufts, James B Robertson, Liara M Gonzalez.

  Intestinal ischemia is a life-threatening emergency with mortality rates of 50-80% due to epithelial cell death and resultant barrier loss.Loss of the epithelial barrier occurs in conditions including intestinal volvulus and neonatal necrotizing enterocolitis.Survival depends on effective epithelial repair; crypt-based intestinal epithelial stem cells (ISCs) are the source of epithelial renewal in homeostasis and after injury. Two ISC populations have been described: 1) active ISC [aISC; highly proliferative; leucine-rich-repeat-containing G-protein-coupled receptor 5 (LGR5+) positive or sex-determining region Y-box 9 -antigen Ki67 positive (SOX9+Ki67+)] and 2) reserve ISC [rISC; less proliferative; homeodomain-only protein X positive (HOPX+)].The contributions of these ISCs have been evaluated both in vivo andin vitrousing a porcine model of mesenteric vascular occlusion to understand mechanisms that modulate ISC recovery responses following ischemic injury. In our previously published work, we observed that rISC conversion to an activated state was associated with decreased HOPXexpression during in vitrorecovery. In the present study, we wished to evaluate the direct role of HOPXon cellular proliferation during recovery after injury. Our data demonstrated that during early in vivo recovery, injury-resistant HOPX+cells maintain quiescence. Subsequent early regeneration within the intestinal crypt occurs around 2 days post injury, a period in which HOPX expression decreased. When HOPX was silenced in vitro,cellular proliferation of injured cells was promoted during recovery. This suggests that HOPXmay serve a functional role in ISC mediated regeneration after injury and could be a target to control ISC proliferation.

Keywords:  HOPX; epithelial repair; ischemia; large animal models; stem cell

DOI:  https://doi.org/10.1152/ajpgi.00165.2021
Nat Metab. 2021 Sep;3(9): 1202-1216

Diet-induced alteration of intestinal stem cell function underlies obesity and prediabetes in mice.

Alexandra Aliluev, Sophie Tritschler, Michael Sterr, Lena Oppenländer, Julia Hinterdobler, Tobias Greisle, Martin Irmler, Johannes Beckers, Na Sun, Axel Walch, Kerstin Stemmer, Alida Kindt, Jan Krumsiek, Matthias H Tschöp, Malte D Luecken, Fabian J Theis, Heiko Lickert, Anika Böttcher.

Excess nutrient uptake and altered hormone secretion in the gut contribute to a systemic energy imbalance, which causes obesity and an increased risk of type 2 diabetes and colorectal cancer. This functional maladaptation is thought to emerge at the level of the intestinal stem cells (ISCs). However, it is not clear how an obesogenic diet affects ISC identity and fate. Here we show that an obesogenic diet induces ISC and progenitor hyperproliferation, enhances ISC differentiation and cell turnover and changes the regional identities of ISCs and enterocytes in mice. Single-cell resolution of the enteroendocrine lineage reveals an increase in progenitors and peptidergic enteroendocrine cell types and a decrease in serotonergic enteroendocrine cell types. Mechanistically, we link increased fatty acid synthesis, Ppar signaling and the Insr-Igf1r-Akt pathway to mucosal changes. This study describes molecular mechanisms of diet-induced intestinal maladaptation that promote obesity and therefore underlie the pathogenesis of the metabolic syndrome and associated complications.

DOI: https://doi.org/10.1038/s42255-021-00458-9