bims-instec 2022-01-09 papers

bims-instec

Biomed News

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

Issue of 2022–01–09
nine papers selected by
Maria-Virginia Giolito, IRFAC/UMR-S1113 INSERM

Enteric glia bring fresh WNT to the intestinal stem cell niche.
YAP1 and PRDM14 converge to promote cell survival and tumorigenesis.
ID2 controls differentiation of enteroendocrine cells in mouse small intestine.
Intestinal organoid co-culture protocol to study cell competition in vitro.
LRP5 promotes cancer stem cell traits and chemoresistance in colorectal cancer.
A 5-FU Precursor Designed to Evade Anabolic and Catabolic Drug Pathways and Activated by Pd Chemistry In Vitro and In Vivo.
Tissue geometry drives deterministic organoid patterning.
The POU2F1-ALDOA axis promotes the proliferation and chemoresistance of colon cancer cells by enhancing glycolysis and the pentose phosphate pathway activity.
Intestine-specific ablation of the Hepatocyte Nuclear Factor 4a (Hnf4a) gene in mice has minimal impact on serum lipids and ileum gene expression profile due to upregulation of its paralog Hnf4g.

Cell Stem Cell. 2022 Jan 06. pii: S1934-5909(21)00487-2. [Epub ahead of print]29(1): 3-4

Enteric glia bring fresh WNT to the intestinal stem cell niche.

Fränze Progatzky, Vassilis Pachnis.

Intestinal stem cells continuously self-renew and differentiate into a variety of specialized epithelial cells that maintain gut health. New research in this issue of Cell Stem Cell (Baghdadi et al., 2022) shows that enteric glial cells regulate the intestinal stem cell niche during regeneration and disease through the production of WNT ligands.

DOI: https://doi.org/10.1016/j.stem.2021.12.003
Dev Cell. 2021 Dec 27. pii: S1534-5807(21)00995-3. [Epub ahead of print]

YAP1 and PRDM14 converge to promote cell survival and tumorigenesis.

Miju Kim, Seav Huong Ly, Yingtian Xie, Gina N Duronio, Dane Ford-Roshon, Justin H Hwang, Rita Sulahian, Jonathan P Rennhack, Jonathan So, Ole Gjoerup, Jessica A Talamas, Maximilien Grandclaudon, Henry W Long, John G Doench, Nilay S Sethi, Marios Giannakis, William C Hahn.

  The transcriptional co-activator YAP1 oncogene is the downstream effector of the Hippo pathway, which regulates tissue homeostasis, organ size, regeneration, and tumorigenesis. Multiple cancers are dependent on sustained expression of YAP1 for cell proliferation, survival, and tumorigenesis, but the molecular basis of this oncogene dependency is not well understood. To identify genes that can functionally substitute for YAP1, we performed a genome-scale genetic rescue screen in YAP1-dependent colon cancer cells expressing an inducible YAP1-specific shRNA. We found that the transcription factor PRDM14 rescued cell proliferation and tumorigenesis upon YAP1 suppression in YAP1-dependent cells, xenografts, and colon cancer organoids. YAP1 and PRDM14 individually activated the transcription of calmodulin 2 (CALM2) and a glucose transporter SLC2A1 upon YAP1 suppression, and CALM2 or SLC2A1 expression was required for the rescue of YAP1 suppression. Together, these findings implicate PRDM14-mediated transcriptional upregulation of CALM2 and SLC2A1 as key components of oncogenic YAP1 signaling and dependency.

Keywords:  Hippo pathway; KRAS; PRDM14; YAP1; colon cancer; oncogene addiction; resistance

DOI:  https://doi.org/10.1016/j.devcel.2021.12.006
Acta Physiol (Oxf). 2022 Jan 05. e13773

ID2 controls differentiation of enteroendocrine cells in mouse small intestine.

Valeriya V Zinina, Frank Ruehle, Patricia Winkler, Lisa Rebmann, Hanna Lukas, Stefanie Möckel, Andreas Diefenbach, Maria Mendez-Lago, Natalia Soshnikova.

   AIMS: The mammalian gut is the largest endocrine organ. Dozens of hormones secreted by enteroendocrine cells regulate a variety of physiological functions of the gut but also of the pancreas and brain. Here, we examined the role of the helix-loop-helix transcription factor ID2 during the differentiation of intestinal stem cells along the enteroendocrine lineage.
METHODS: To assess the functions of ID2 in the adult mouse small intestine, we used single-cell RNA sequencing, genetically modified mice, and organoid assays.
RESULTS: We found that in the adult intestinal epithelium Id2 is predominantly expressed in enterochromaffin and peptidergic enteroendocrine cells. Consistently, the loss of Id2 leads to the reduction of Chromogranin A-positive enteroendocrine cells. In contrast, the numbers of tuft cells are increased in Id2 mutant small intestine. Moreover, ablation of Id2 elevates the numbers of Serotonin+ enterochromaffin cells and Ghrelin+ X-cells in the posterior part of the small intestine. Finally, ID2 acts downstream of BMP signalling during the differentiation of Glucagon Like Peptide-1+ L-cells and Cholecystokinin+ I-cells towards Neurotensin+ PYY+ N-cells.
CONCLUSION: ID2 plays an important role in cell fate decisions in the adult small intestine. Firstly, ID2 is essential for establishing a differentiation gradient for enterochromaffin and X-cells along the anterior-posterior axis of the gut. Next, ID2 is necessary for the differentiation of N-cells thus ensuring a differentiation gradient along the crypt-villi axis. Finally, ID2 suppresses the commitment of secretory intestinal epithelial progenitors towards tuft cell lineage and thus controls host immune response to commensal and parasitic microbiota.

Keywords:  endocrine cell; gut; hormone; ID2; organoid; single-cell RNA-sequencing; tuft cell

DOI:  https://doi.org/10.1111/apha.13773
STAR Protoc. 2022 Mar 18. 3(1): 101050

Intestinal organoid co-culture protocol to study cell competition in vitro.

Sanne M van Neerven, Rana Ramadan, Milou S van Driel, David J Huels, Louis Vermeulen.

  Intestinal organoid cultures are a powerful tool to study epithelial cells in vitro, as they are able to proliferate and differentiate into all cell lineages observed in vivo. Co-culturing organoids with distinct genetic backgrounds provides an excellent approach to study contact dependent and independent interactions between healthy and mutant epithelial intestinal cells. Here, we provide 2D and 3D approaches to mouse organoid co-cultures using fluorescently labeled organoids and demonstrate the analysis of these co-cultures using flow cytometry and microscopy-based approaches. For complete details on the use and execution of this profile, please refer to van Neerven et al., 2021.

Keywords:  Cancer; Cell Biology; Cell culture; Cell-based Assays; Flow Cytometry/Mass Cytometry; Microscopy; Organoids

DOI:  https://doi.org/10.1016/j.xpro.2021.101050
J Cell Mol Med. 2022 Jan 07.

LRP5 promotes cancer stem cell traits and chemoresistance in colorectal cancer.

Xiaobo Nie, Huiyang Liu, Wenling Ye, Xiaoyun Wei, Lili Fan, Han Ma, Lanqing Li, Wanting Xue, Wenting Qi, Yan-Dong Wang, Wei-Dong Chen.

  The overactivation of canonical Wnt/β-catenin pathway and the maintenance of cancer stem cells (CSCs) are essential for the onset and malignant progression of most human cancers. However, their regulatory mechanism in colorectal cancer (CRC) has not yet been well demonstrated. Low-density lipoprotein receptor-related protein 5 (LRP5) has been identified as an indispensable co-receptor with frizzled family members for the canonical Wnt/β-catenin signal transduction. Herein, we show that activation of LRP5 gene promotes CSCs-like phenotypes, including tumorigenicity and drug resistance in CRC cells, through activating the canonical Wnt/β-catenin and IL-6/STAT3 signalling pathways. Clinically, the expression of LRP5 is upregulated in human CRC tissues and closely associated with clinical stages of patients with CRC. Further analysis showed silencing of endogenous LRP5 gene is sufficient to suppress the CSCs-like phenotypes of CRC through inhibiting these two pathways. In conclusion, our findings not only reveal a regulatory cross-talk between canonical Wnt/β-catenin signalling pathway, IL-6/STAT3 signalling pathway and CD133-related stemness that promote the malignant behaviour of CRC, but also provide a valuable target for the diagnosis and treatment of CRC.

Keywords:  IL-6/STAT3 signalling pathway; LRP5; cancer stem cells; canonical Wnt/β-catenin signalling pathway; colorectal cancer

DOI:  https://doi.org/10.1111/jcmm.17164
J Med Chem. 2022 Jan 03.

A 5-FU Precursor Designed to Evade Anabolic and Catabolic Drug Pathways and Activated by Pd Chemistry In Vitro and In Vivo.

Catherine Adam, Thomas L Bray, Ana M Pérez-López, Ee Hong Tan, Belén Rubio-Ruiz, Daniel J Baillache, Douglas R Houston, Mark J Salji, Hing Y Leung, Asier Unciti-Broceta.

5-Fluorouracil (5-FU) is an antineoplastic antimetabolite that is widely administered to cancer patients by bolus injection, especially to those suffering from colorectal and pancreatic cancer. Because of its suboptimal route of administration and dose-limiting toxicities, diverse 5-FU prodrugs have been developed to confer oral bioavailability and increase the safety profile of 5-FU chemotherapy regimens. Our contribution to this goal is presented herein with the development of a novel palladium-activated prodrug designed to evade the metabolic machinery responsible for 5-FU anabolic activation and catabolic processing. The new prodrug is completely innocuous to cells and highly resistant to metabolization by primary hepatocytes and liver S9 fractions (the main metabolic route for 5-FU degradation), whereas it is rapidly converted into 5-FU in the presence of a palladium (Pd) source. In vivo pharmokinetic analysis shows the prodrug is rapidly and completely absorbed after oral administration and exhibits a longer half-life than 5-FU. In vivo efficacy studies in a xenograft colon cancer model served to prove, for the first time, that orally administered prodrugs can be locally converted to active drugs by intratumorally inserted Pd implants.

DOI: https://doi.org/10.1021/acs.jmedchem.1c01733
Science. 2022 Jan 07. 375(6576): eaaw9021

Tissue geometry drives deterministic organoid patterning.

N Gjorevski, M Nikolaev, T E Brown, O Mitrofanova, N Brandenberg, F W DelRio, F M Yavitt, P Liberali, K S Anseth, M P Lutolf.

[Figure: see text].

DOI: https://doi.org/10.1126/science.aaw9021
Oncogene. 2022 Jan 08.

The POU2F1-ALDOA axis promotes the proliferation and chemoresistance of colon cancer cells by enhancing glycolysis and the pentose phosphate pathway activity.

Jinguan Lin, Longzheng Xia, Linda Oyang, Jiaxin Liang, Shiming Tan, Nayiyuan Wu, Pin Yi, Qing Pan, Shan Rao, Yaqian Han, Yanyan Tang, Min Su, Xia Luo, Yiqing Yang, Xiaohui Chen, Lixia Yang, Yujuan Zhou, Qianjin Liao.

Cancer metabolic reprogramming enhances its malignant behaviors and drug resistance, which is regulated by POU domain transcription factors. This study explored the effect of POU domain class 2 transcription factor 1 (POU2F1) on metabolic reprogramming in colon cancer. The POU2F1 expression was analyzed in GEO dataset, TCGA cohorts and human colon cancer tissues by bioinformatics and immunohistochemistry. The effects of altered POU2F1 expression on proliferation, glucose metabolism and oxaliplatin sensitivity of colon cancer cells were tested. The impacts of POU2F1 on aldolase A (ALDOA) expression and malignant behaviors of colon cancer cells were examined. We found that up-regulated POU2F1 expression was associated with worse prognosis and oxaliplatin resistance in colon cancer. POU2F1 enhanced the proliferation, aerobic glycolysis and the pentose phosphate pathway (PPP) activity, but reduced oxidative stress and apoptosis in colon cancer cells, dependent on up-regulating ALDOA expression. Mechanistically, POU2F1 directly bound to the ALDOA promoter to enhance the ALDOA promoter activity in colon cancer cells. Moreover, activation of the POU2F1-ALDOA axis decreased the sensitivity to oxaliplatin in colon cancer cells. These data indicate that the POU2F1-ALDOA axis promotes the progression and oxaliplatin resistance by enhancing metabolic reprogramming in colon cancer. Our findings suggest that the POU2F1-ALDOA axis may be new therapeutic targets to overcome oxaliplatin resistance in colon cancer.

DOI: https://doi.org/10.1038/s41388-021-02148-y
Biochim Biophys Acta Mol Cell Biol Lipids. 2021 Dec 29. pii: S1388-1981(21)00236-5. [Epub ahead of print] 159108

Intestine-specific ablation of the Hepatocyte Nuclear Factor 4a (Hnf4a) gene in mice has minimal impact on serum lipids and ileum gene expression profile due to upregulation of its paralog Hnf4g.

Efstathia Thymiakou, Efsevia Xenikaki, Dimitris Kardassis.

  Ablation of the gene encoding the nuclear receptor Hepatocyte Nuclear Factor 4a (Hnf4a) in the liver strongly affects HDL concentration, structure and functionality but the role of this receptor in the intestine, the second organ contributing to serum HDL levels, has been overlooked. In the present study we show that mice with intestine-specific ablation of Hnf4a (H4IntKO) had undetectable levels of ΗΝF4A in ileum, proximal and distal colon but normal expression in liver. H4IntKO mice presented normal serum lipid levels, HDL-C and particle size (α1-α3). The expression of the major HDL biogenesis genes Apoa1, Abca1, Lcat was not affected but there was significant increase in Apoc3 as well as in Hnf4g, a paralog of Hnf4a. RNA-sequencing identified metabolic pathways significantly affected by Hnf4a ablation such as type II diabetes, glycolysis, gluconeogenesis and p53 signaling. Chromatin immunoprecipitation assays showed that HNF4G bound to various apolipoprotein gene promoters in control mice but its binding affinity was reduced in the ileum of H4IntKO mice suggesting a redundancy but also a cooperation between the two factors. In the distal colon of H4IntKO mice, where both HNF4A and HNF4G are absent and in a mouse model of DSS-induced colitis presenting decreased levels of HNF4A, most lipoprotein genes were strongly downregulated. In conclusion, Hnf4a ablation in mice does not significantly affect serum lipid levels or lipoprotein gene expression in ileum possibly due to compensatory effects by its paralog Hnf4g in this tissue.

Keywords:  Colitis; Gene ablation; Hepatocyte Nuclear Factor 4; High density lipoproteins; Ileum; Intestine-specific; Mice; RNA-seq

DOI:  https://doi.org/10.1016/j.bbalip.2021.159108