bims-instec 2022-12-11 papers

bims-instec

Biomed News

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

Issue of 2022–12–11
23 papers selected by
Maria-Virginia Giolito, Free University of Brussels

HNF4α acts as upstream functional regulator of intestinal Wnt3 and Paneth cell fate.
Epithelial TGFβ engages growth-factor signalling to circumvent apoptosis and drive intestinal tumourigenesis with aggressive features.
Gut microbiota promotes stem cell differentiation through macrophage and mesenchymal niches in early postnatal development.
Yap governs a lineage-specific neuregulin1 pathway-driven adaptive resistance to RAF kinase inhibitors.
Editorial: Recent advances of drug resistance research in colorectal cancer therapy.
Editorial: Non-genetic adaptive drug resistance in cancer.
Single-cell transcriptome analysis of tumor immune microenvironment characteristics in colorectal cancer liver metastasis.
The use of immunotherapy in metastatic microsatellite-stable colorectal cancer.
Tadalafil increases the antitumor activity of 5-FU through inhibiting PRMT5-mediated glycolysis and cell proliferation in colorectal cancer.
A Human Vascularized Micro-Tumor Model of Patient-Derived Colorectal Cancer Recapitulates Clinical Disease.
Role of NRF2 in Colorectal Cancer Prevention and Treatment.
Intestinal models based on biomimetic scaffolds with an ECM micro-architecture and intestinal macro-elasticity: close to intestinal tissue and immune response analysis.
Low Expression of AGPAT5 Is Associated With Clinical Stage and Poor Prognosis in Colorectal Cancer and Contributes to Tumour Progression.
VASN promotes colorectal cancer progression by activating the YAP/TAZ and AKT signaling pathways via YAP.
RNF43 induces the turnover of protease-activated receptor 2 in colon cancer.
Current concepts of anti-EGFR targeting in metastatic colorectal cancer.
Targeting ferroptosis to treat colorectal cancer.
A first-in-human study of the anti-LAG-3 antibody favezelimab plus pembrolizumab in previously treated, advanced microsatellite stable colorectal cancer.
An integrated map of fibroblastic populations in human colon mucosa and cancer tissues.
Predictive biomarkers of colon cancer immunotherapy: Present and future.
Low intensity treatment in metastatic colorectal cancer.
Spatial transcriptomics add a new dimension to our understanding of the gut.
Cytokine-mediated crosstalk between cancer stem cells and their inflammatory niche from the colorectal precancerous adenoma stage to the cancerous stage: Mechanisms and clinical implications.

Cell Mol Gastroenterol Hepatol. 2022 Dec 01. pii: S2352-345X(22)00246-6. [Epub ahead of print]

HNF4α acts as upstream functional regulator of intestinal Wnt3 and Paneth cell fate.

Christine Jones, Mariano Avino, Véronique Giroux, Francois Boudreau.

   BACKGROUND AND AIMS: The intestinal epithelium intrinsically renews itself ex vivo via the proliferation of Lgr5+ intestinal stem cells, which is sustained by the establishment of an epithelial stem cell niche. Differentiated Paneth cells are the main source of epithelial-derived niche factor supplies, and produce Wnt3 as an essential factor in supporting Lgr5+ stem cell activity in the absence of redundant mesenchymal Wnts. Maturation of Paneth cells depends on canonical Wnt signaling, but few transcriptional regulators have been identified to this end. The role of HNF4α in intestinal epithelial cell differentiation is considered redundant with its paralog, HNF4γ. However, it is unclear whether HNF4α alone controls intrinsic intestinal epithelial cell growth and fate in the absence of a mesenchymal niche.
METHODS: We used transcriptomic analyses to dissect the role of HNF4α in the maintenance of jejunal epithelial culture when cultured ex vivo as enteroids, in the presence or absence of compensatory mesenchymal cells.
RESULTS: HNF4α plays a crucial role in supporting the growth and survival of jejunal enteroids. Transcriptomic analyses revealed an autonomous function of HNF4α in Wnt3 transcriptional regulation and Paneth cell differentiation. We showed that Wnt3a supplementation or co-culture with intestinal sub-epithelial mesenchymal cells reversed cell death and transcriptional changes caused by the deletion of Hnf4a in jejunal enteroids.
CONCLUSIONS: Our results support the intrinsic epithelial role of HNF4α in regulating Paneth cell homeostasis and intestinal epithelium renewal in the absence of compensatory Wnt signaling.

Keywords:  Enteroids; HNF4α; Paneth Cells; Wnt3

DOI:  https://doi.org/10.1016/j.jcmgh.2022.11.010
Nat Commun. 2022 Dec 07. 13(1): 7551

Epithelial TGFβ engages growth-factor signalling to circumvent apoptosis and drive intestinal tumourigenesis with aggressive features.

Dustin J Flanagan, Raheleh Amirkhah, David F Vincent, Nuray Gundaz, Pauline Gentaz, Patrizia Cammareri, Aoife J McCooey, Amy M B McCorry, Natalie C Fisher, Hayley L Davis, Rachel A Ridgway, Jeroen Lohuis, Joshua D G Leach, Rene Jackstadt, Kathryn Gilroy, Elisa Mariella, Colin Nixon, William Clark, Ann Hedley, Elke K Markert, Douglas Strathdee, Laurent Bartholin, Keara L Redmond, Emma M Kerr, Daniel B Longley, Fiona Ginty, Sanghee Cho, Helen G Coleman, Maurice B Loughrey, Alberto Bardelli, Timothy S Maughan, Andrew D Campbell, Mark Lawler, Simon J Leedham, Simon T Barry, Gareth J Inman, Jacco van Rheenen, Philip D Dunne, Owen J Sansom.

The pro-tumourigenic role of epithelial TGFβ signalling in colorectal cancer (CRC) is controversial. Here, we identify a cohort of born to be bad early-stage (T1) colorectal tumours, with aggressive features and a propensity to disseminate early, that are characterised by high epithelial cell-intrinsic TGFβ signalling. In the presence of concurrent Apc and Kras mutations, activation of epithelial TGFβ signalling rampantly accelerates tumourigenesis and share transcriptional signatures with those of the born to be bad T1 human tumours and predicts recurrence in stage II CRC. Mechanistically, epithelial TGFβ signalling induces a growth-promoting EGFR-signalling module that synergises with mutant APC and KRAS to drive MAPK signalling that re-sensitise tumour cells to MEK and/or EGFR inhibitors. Together, we identify epithelial TGFβ signalling both as a determinant of early dissemination and a potential therapeutic vulnerability of CRC's with born to be bad traits.

DOI: https://doi.org/10.1038/s41467-022-35134-3
Immunity. 2022 Nov 29. pii: S1074-7613(22)00593-3. [Epub ahead of print]

Gut microbiota promotes stem cell differentiation through macrophage and mesenchymal niches in early postnatal development.

Ji-Eun Kim, Bo Li, Lijiang Fei, Rachael Horne, Dorothy Lee, Adrian Kwan Loe, Hiromu Miyake, Eda Ayar, Dae-Kyum Kim, Michael G Surette, Dana J Philpott, Philip Sherman, Guoji Guo, Agostino Pierro, Tae-Hee Kim.

  Intestinal stem cell maturation and development coincide with gut microbiota exposure after birth. Here, we investigated how early life microbial exposure, and disruption of this process, impacts the intestinal stem cell niche and development. Single-cell transcriptional analysis revealed impaired stem cell differentiation into Paneth cells and macrophage specification upon antibiotic treatment in early life. Mouse genetic and organoid co-culture experiments demonstrated that a CD206+ subset of intestinal macrophages secreted Wnt ligands, which maintained the mesenchymal niche cells important for Paneth cell differentiation. Antibiotics and reduced numbers of Paneth cells are associated with the deadly infant disease, necrotizing enterocolitis (NEC). We showed that colonization with Lactobacillus or transfer of CD206+ macrophages promoted Paneth cell differentiation and reduced NEC severity. Together, our work defines the gut microbiota-mediated regulation of stem cell niches during early postnatal development.

Keywords:  Paneth cells; Wnt signaling; antibiotics; intestine; macrophages; microbiota; necrotizing enterocolitis; stem cell niche

DOI:  https://doi.org/10.1016/j.immuni.2022.11.003
Mol Cancer. 2022 Dec 07. 21(1): 213

Yap governs a lineage-specific neuregulin1 pathway-driven adaptive resistance to RAF kinase inhibitors.

Maria E R Garcia-Rendueles, Gnana Krishnamoorthy, Mahesh Saqcena, Adrian Acuña-Ruiz, Giovanna Revilla, Elisa de Stanchina, Jeffrey A Knauf, Rona Lester, Bin Xu, Ronald A Ghossein, James A Fagin.

   BACKGROUND: Inactivation of the Hippo pathway promotes Yap nuclear translocation, enabling execution of a transcriptional program that induces tissue growth. Genetic lesions of Hippo intermediates only identify a minority of cancers with illegitimate YAP activation. Yap has been implicated in resistance to targeted therapies, but the mechanisms by which YAP may impact adaptive resistance to MAPK inhibitors are unknown.
METHODS: We screened 52 thyroid cancer cell lines for illegitimate nuclear YAP localization by immunofluorescence and fractionation of cell lysates. We engineered a doxycycline (dox)-inducible thyroid-specific mouse model expressing constitutively nuclear YAPS127A, alone or in combination with endogenous expression of either HrasG12V or BrafV600E. We also generated cell lines expressing dox-inducible sh-miR-E-YAP and/or YAPS127A. We used cell viability, invasion assays, immunofluorescence, Western blotting, qRT-PCRs, flow cytometry and cell sorting, high-throughput bulk RNA sequencing and in vivo tumorigenesis to investigate YAP dependency and response of BRAF-mutant cells to vemurafenib.
RESULTS: We found that 27/52 thyroid cancer cell lines had constitutively aberrant YAP nuclear localization when cultured at high density (NU-YAP), which rendered them dependent on YAP for viability, invasiveness and sensitivity to the YAP-TEAD complex inhibitor verteporfin, whereas cells with confluency-driven nuclear exclusion of YAP (CYT-YAP) were not. Treatment of BRAF-mutant thyroid cancer cells with RAF kinase inhibitors resulted in YAP nuclear translocation and activation of its transcriptional output. Resistance to vemurafenib in BRAF-mutant thyroid cells was driven by YAP-dependent NRG1, HER2 and HER3 activation across all isogenic human and mouse thyroid cell lines tested, which was abrogated by silencing YAP and relieved by pan-HER kinase inhibitors. YAP activation induced analogous changes in BRAF melanoma, but not colorectal cells.
CONCLUSIONS: YAP activation in thyroid cancer generates a dependency on this transcription factor. YAP governs adaptive resistance to RAF kinase inhibitors and induces a gene expression program in BRAFV600E-mutant cells encompassing effectors in the NRG1 signaling pathway, which play a central role in the insensitivity to MAPK inhibitors in a lineage-dependent manner. HIPPO pathway inactivation serves as a lineage-dependent rheostat controlling the magnitude of the adaptive relief of feedback responses to MAPK inhibitors in BRAF-V600E cancers.

Keywords:  BRAF; Dependency; HER2; HER3; Hippo; Lapatinib; NRG1; Resistance; Thyroid cancer; Vemurafenib; YAP

DOI:  https://doi.org/10.1186/s12943-022-01676-9
Front Oncol. 2022 ;12 1059223

Editorial: Recent advances of drug resistance research in colorectal cancer therapy.

Chenyang Ye, Yanhong Deng, Weijing Sun, Ying Yuan.



Keywords:  cancer treatment; colorectal cancer; drug resistance (DR); drug resistance mechanism; prognosis analysis

DOI:  https://doi.org/10.3389/fonc.2022.1059223
Front Cell Dev Biol. 2022 ;10 998339

Editorial: Non-genetic adaptive drug resistance in cancer.

Michaël Cerezo, Xiaoxiao Sun, Shensi Shen.



Keywords:  cellular plasticity; drug resistance; non-genetic adaptation; persistent cancer cells; therapeutic targets; tumor relapse

DOI:  https://doi.org/10.3389/fcell.2022.998339
Ann Transl Med. 2022 Nov;10(21): 1170

Single-cell transcriptome analysis of tumor immune microenvironment characteristics in colorectal cancer liver metastasis.

Yiting Geng, Jun Feng, Hao Huang, Ying Wang, Xing Yi, Shanshan Wei, Mingyue Zhang, Zhong Li, Wei Wang, Wenwei Hu.

   Background: Liver metastasis is the leading cause of death in colorectal cancer (CRC) patients, and the precise mechanisms remain unclear. In this study, single-cell RNA sequencing (scRNA-seq) was used to analyze the cellular and molecular heterogeneity between CRC primary lesion and corresponding liver metastasis, and to clarify the characteristics of the tumor microenvironment (TME) in synchronous liver metastasis of CRC.
Methods: A case of microsatellite stable (MSS) sigmoid carcinoma with synchronous liver metastasis was selected, and tissues from the primary tumor and the liver metastasis were collected for scRNA-seq. The EdgeR package software was used to identify the differentially expressed genes between cells. Gene Set Enrichment Analysis (GSEA) was performed and the clusterProfiler R package was used for Gene Ontology (GO) enrichment analysis. The SCENIC and CellphoneDB packages were used to reconstruct the transcriptional regulatory networks and to analyze the intercellular interaction network, respectively.
Results: Compared to the primary tumor, the proportion of myeloid cells in the metastatic tumor was significantly increased, while B cells and plasma cells were decreased. In the metastatic tumor, the myeloid-derived suppressor cell (MDSC) characteristic gene, mannose receptor C-type 1 (MRC1) and tumor associated macrophage 2 (TAM2)-related gene, were highly expressed. Furthermore, angiogenesis, oxidative phosphorylation, and endothelial mesenchymal transition (EMT) of myeloid cells were also significantly enhanced. There were less myeloid cells in primary tumors, and these were mainly monocytes and TAM1; while the number of TAM2 was significantly upregulated in the metastatic samples. In liver metastasis, the T cell population was exhausted, and this was accompanied by a significant increase in the number of CD4+ T cells and a decrease in the number of CD8+ T cells. Furthermore, some immune checkpoint molecules were highly expressed. Interactions between myeloid cells and other cell populations appeared to be strong.
Conclusions: The TME of CRC liver metastasis is significantly immunosuppressed. Interactions between myeloid cells and other cell populations in the TME contribute to the establishment of a pro-metastatic niche that promotes colonization and growth of CRC cells in the liver. TAMs may be a potential immunotherapeutic target for MSS CRC.

Keywords:  Colorectal cancer (CRC); liver metastasis; single-cell RNA sequencing; tumor immune microenvironment; tumor microenvironment

DOI:  https://doi.org/10.21037/atm-22-5270
Clin Adv Hematol Oncol. 2022 Dec;20(12): 703-704

The use of immunotherapy in metastatic microsatellite-stable colorectal cancer.

Christopher Lieu.
Cancer Metab. 2022 Dec 06. 10(1): 22

Tadalafil increases the antitumor activity of 5-FU through inhibiting PRMT5-mediated glycolysis and cell proliferation in colorectal cancer.

Yao Shen, Pan Zhao, Kewei Dong, Jiajia Wang, Huichen Li, Mengyang Li, Ruikai Li, Suning Chen, Yuxia Shen, Zhiyu Liu, Mianjiao Xie, Peng Shen, Jian Zhang.

   BACKGROUND: Protein arginine methyltransferase 5 (PRMT5) is upregulated in multiple tumors and plays a pivotal role in cancer cell proliferation. However, the role of PRMT5 in colorectal cancer remains poorly understood.
METHODS: We detected the expression level of PRMT5 and glycolytic enzymes using online databases and colorectal cancer cell lines by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR), and western blotting. And MTT and colony formation assays were conducted to investigate cell proliferation. Then, we evaluated ECAR and OCR levels using a biological energy analyzer to investigate the energy status of colorectal cancer, and the transcriptional regulation was detected by dual luciferase reporter assay and ChIP assay. Finally, the efficacy of combined treatment of tadalafil and 5-FU was verified.
RESULTS: PRMT5 was highly expressed in colorectal cancer tissues compared with their normal counterparts and correlated with poor prognosis in CRC patients. Then, we demonstrated that PRMT5 knockdown or loss of function attenuated the viability of CRC cells, while overexpression of PRMT5 promoted cell proliferation. Mechanistically, PRMT5 enhanced glycolysis through transcriptionally activating LDHA expression. In addition, the PRMT5 inhibitor, tadalafil, rendered CRC cells sensitive to antitumor agent 5-FU in vitro and in vivo.
CONCLUSIONS: Our data indicates that PRMT5 promoted colorectal cancer proliferation partially through activating glycolysis and may be a potential target for colorectal cancer therapy.

Keywords:  5-FU; Colorectal cancer; PRMT5; Tadalafil

DOI:  https://doi.org/10.1186/s40170-022-00299-4
Transl Res. 2022 Dec 05. pii: S1931-5244(22)00278-X. [Epub ahead of print]

A Human Vascularized Micro-Tumor Model of Patient-Derived Colorectal Cancer Recapitulates Clinical Disease.

Stephanie J Hachey, Agua Sobrino, John G Lee, Mehraneh D Jafari, Samuel J Klempner, Eric J Puttock, Robert A Edwards, John S Lowengrub, Marian L Waterman, Jason A Zell, Christopher C W Hughes.

  Accurately modeling tumor biology and testing novel therapies on patient-derived cells is critically important to developing therapeutic regimens personalized to a patient's specific disease. The vascularized micro-tumor (VMT), or "tumor-on-a-chip", is a physiologic preclinical cancer model that incorporates key features of the native human tumor microenvironment within a transparent microfluidic platform, allowing rapid drug screening in vitro. Herein we optimize methods for generating patient-derived VMT (pVMT) using fresh colorectal cancer (CRC) biopsies and surgical resections to test drug sensitivities at the individual patient level. In response to standard chemotherapy and TGF-βR1 inhibition, we observe heterogeneous responses between pVMT derived from 6 patient biopsies, with the pVMT recapitulating tumor growth, histological features, metabolic heterogeneity and drug responses of actual CRC tumors. Our results suggest that a translational infrastructure providing rapid information from patient-derived tumor cells in the pVMT, as established in this study, will support efforts to improve patient outcomes.

Keywords:  microphysiologic | oncology | personalized medicine | disease modeling

DOI:  https://doi.org/10.1016/j.trsl.2022.11.011
Technol Cancer Res Treat. 2022 Jan-Dec;21:21 15330338221105736

Role of NRF2 in Colorectal Cancer Prevention and Treatment.

Jiaxiang Li, Dan Wang, Yifei Liu, Yanhong Zhou.

  Nuclear factor erythroid 2-related factor 2 (NRF2) is a basic leucine zipper protein that participates in a complex regulatory network in the body. The activation of NRF2 can prevent and treat colorectal cancer (CRC). A variety of natural compounds can activate NRF2 to inhibit oxidative stress and inflammation to prevent the occurrence and development of CRC, inhibit the proliferation of CRC cells and induce their apoptosis. However, some studies have also shown that it also has negative effects on CRC, such as overexpression of NRF2 can promote the growth of colorectal tumors and increase the drug resistance of chemotherapeutic drugs such as 5-fluorouracil and oxaliplatin. Therefore, inhibition of NRF2 can also be helpful in the treatment of CRC. In this study, we analyze the current research progress of NRF2 in CRC from various aspects to provide new ideas for prevention and treatment based on the NRF2 signaling pathway in CRC.

Keywords:  colorectal cancer; inflammatory-associated colorectal cancer; nuclear factor E2-related factor 2; prevention; treatment

DOI:  https://doi.org/10.1177/15330338221105736
Biomater Sci. 2022 Dec 07.

Intestinal models based on biomimetic scaffolds with an ECM micro-architecture and intestinal macro-elasticity: close to intestinal tissue and immune response analysis.

Yue Li, Meng-Xin Hu, Ming Yan, Ya-Xin Guo, Xue-Ke Ma, Jian-Zhong Han, Yu-Mei Qin.

The synergetic biological effect of scaffolds with biomimetic properties including the ECM micro-architecture and intestinal macro-mechanical properties on intestinal models in vitro remains unclear. Here, we investigate the profitable role of biomimetic scaffolds on 3D intestinal epithelium models. Gelatin/bacterial cellulose nanofiber composite scaffolds crosslinked by the Maillard reaction are tuned to mimic the chemical component, nanofibrous network, and crypt architecture of intestinal ECM collagen and the stability and mechanical properties of intestinal tissue. In particular, scaffolds with comparable elasticity and viscoelasticity of intestinal tissue possess the highest biocompatibility and best cell proliferation and differentiation ability, which makes the intestinal epithelium models closest to their counterpart intestinal tissues. The constructed duodenal epithelium models and colon epithelium models are utilized to assess the immunobiotics-host interactions, and both of them can sensitively respond to foreign microorganisms, but the secretion levels of cytokines are intestinal cell specific. The results demonstrate that probiotics alleviate the inflammation and cell apoptosis induced by Escherichia coli, indicating that probiotics can protect the intestinal epithelium from damage by inhibiting the adhesion and invasion of E. coli to intestinal cells. The designed biomimetic scaffolds can serve as powerful tools to construct in vitro intestinal epithelium models, providing a convenient platform to screen intestinal anti-inflammatory components and even to assess other physiological functions of the intestine.

DOI: https://doi.org/10.1039/d2bm01051h
Clin Med Insights Oncol. 2022 ;16 11795549221137399

Low Expression of AGPAT5 Is Associated With Clinical Stage and Poor Prognosis in Colorectal Cancer and Contributes to Tumour Progression.

Jia Zang, Juanjuan Sun, WenChao Xiu, Xiaoshuang Liu, Yunsheng Chai, Yanyan Zhou.

   Background: Colorectal cancer (CRC) has a high prevalence and poor prognosis. This study aimed to identify biomarkers related to the clinical stage (I-IV) of CRC.
Methods: The LinkedOmics database was used as the discovery cohort, and two Gene Expression Omnibus (GEO) databases (GSE41258 and GSE422848) served as validation cohorts. The trend test of genes related to clinical stage (I-IV) of CRC patients was identified by the Jonckheere-Terpstra test. The cBioPortal database, Gene Expression Profiling Interactive Analysis (GEPIA) and PrognoScan databases were used to explore the expression change and prognostic value of clinical stage-related genes in CRC patients. CRC cells overexpressed AGPAT5 were constructed and used for cell counting kit-8 (CCK-8), flow cytometric, and wound healing assays in vitro.
Results: We identified four clinical stage-related genes, GSR, AGPAT5, CRLF1, and NPR3, in CRC. The CNA frequencies of GSR, CRLF1, AGPAT5, and NPR3 occurred in 11%, 2.4%, 13%, and 3% of patients, respectively. The expression of GSR and AGPAT5 tended to decrease with CRC stage (I-IV) progression, and the expression of CRLF1 and NPR3 tended to increase with CRC stage (I-IV) progression. Compared with the normal group, AGPAT5 expression was markedly decreased in stage IV CRC. Higher GSR and AGPAT5 expression levels were associated with better overall survival (OS) and disease-free survival (DFS) in CRC patients. Lower CRLF1 and NPR3 expression levels were associated with better OS and DFS in CRC. GSR, CRLF1, AGPAT5, and NPR3 expression were related to CRC progression, microsatellite instability, and tumour purity in CRC. Furthermore, AGPAT5 was downregulated in CRC cell lines, and overexpression of AGPAT5 inhibited cell proliferation and migration and promoted cell apoptosis in CRC cells.
Conclusion: Low AGPAT5 expression may serve as a poor prognostic factor and clinical stage biomarker in CRC. In addition, AGPAT5 acts as a tumour suppressor in CRC progression.

Keywords:  AGPAT5; Colorectal cancer; biomarkers; clinical stage; prognosis; tumour suppressor

DOI:  https://doi.org/10.1177/11795549221137399
FASEB J. 2023 Jan;37(1): e22688

VASN promotes colorectal cancer progression by activating the YAP/TAZ and AKT signaling pathways via YAP.

Weiye Liang, Jia Zuo, Mingkai Liu, Yuling Su, Baoyin Guo, Jiangtao Hou, Qi Xing, Yinglong Peng, Lian Fang, Yihui Cao, Jiajie Shan, Ruixia Sun, Jie Zhao, Jian Wang.

  Colorectal cancer (CRC) is one of the most common gastrointestinal malignancies. Vasorin (VASN) has been reported to be critical in tumor development and angiogenesis. However, VASN has not been reported in CRC, and its role is unclear. In this study, VASN expression is upregulated in CRC compared with the normal tissues, and VASN expression positively correlates with N stage and poor overall survival by analysis of different datasets and 32 CRC clinicopathologic samples. Overexpression of VASN significantly promotes CRC cell progression, including proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT), while knockdown of VASN inhibits CRC progression. We found that VASN was associated with the YAP/TAZ and PI3K/AKT pathways by gene set enrichment analysis (GSEA) and gene ontology (GO) analysis. Notably, western blotting, immunofluorescence staining and co-immunofluorescence (co-IP) confirmed that VASN could interact with YAP and activate the YAP/TAZ and PTEN/PI3K/AKT pathways, and knockdown of YAP reversed this effect. Importantly, our findings indicate that VASN interacts with YAP to inhibit YAP phosphorylation and stimulates CRC proliferation, migration, and invasion through activation of the YAP/TAZ-TEAD target gene CTGF and PTEN/PI3K/AKT pathways. Our results also show that knockdown of YAP reverses the cellular phenotype induced by increased VASN. In conclusion, our study reveals that VASN acts as an oncogene to stimulate tumor progression in CRC, providing new insights into the molecular mechanisms of CRC development and representing a possible novel biomarker for CRC.

Keywords:  AKT; TAZ; VASN; YAP; colorectal cancer; invasion; migration; proliferation

DOI:  https://doi.org/10.1096/fj.202201181R
FASEB J. 2023 Jan;37(1): e22675

RNF43 induces the turnover of protease-activated receptor 2 in colon cancer.

Jeetendra Kumar Nag, Priyanga Appasamy, Shoshana Sedley, Hodaya Malka, Tatyana Rudina, Rachel Bar-Shavit.

  Post-translational modification of G-protein coupled receptors (GPCRs) plays a central role in tissue hemostasis and cancer. The molecular mechanism of post-translational regulation of protease-activated receptors (PARs), a subgroup of GPCRs is yet understudied. Here we show that the cell-surface transmembrane E3 ubiquitin ligase ring finger 43 (RNF43) is a negative feedback regulator of PAR2 , impacting PAR2 -induced signaling and colon cancer growth. RNF43 co-associates with PAR2 , promoting its membrane elimination and degradation as shown by reduced cell surface biotinylated PAR2 levels and polyubiquitination. PAR2 degradation is rescued by R-spondin2 in the presence of leucine-rich repeat-containing G-protein-coupled receptor5 (LGR5). In fact, PAR2 acts jointly with LGR5, as recapitulated by increased β-catenin levels, transcriptional activity, phospho-LRP6, and anchorage-independent colony growth in agar. Animal models of the chemically induced AOM/DSS colon cancer of wt versus Par2/f2rl1 KO mice as also the 'spleen-liver' colon cancer metastasis, allocated a central role for PAR2 in colon cancer growth and development. RNF43 is abundantly expressed in the Par2/f2rl1 KO-treated AOM/DSS colon tissues while its level is very low to nearly null in colon cancer adenocarcinomas of the wt mice. The same result is obtained in the 'spleen-liver' model of spleen-inoculated cells, metastasized to the liver. High RNF43 expression is observed in the liver upon shRNA -Par2 silencing. "Limited-dilution-assay" performed in mice in-vivo, assigned PAR2 as a member of the cancer stem cell niche compartment. Collectively, we elucidate an original regulation of PAR2 oncogene, a member of cancer stem cells, by RNF43 ubiquitin ligase. It impacts β-catenin signaling and colon cancer growth.

Keywords:  GPCRs; R-spondin; RNF43; colon cancer; protease-activated receptors (PARs); β-catenin

DOI:  https://doi.org/10.1096/fj.202200858RR
Front Oncol. 2022 ;12 1048166

Current concepts of anti-EGFR targeting in metastatic colorectal cancer.

Bernhard Doleschal, Andreas Petzer, Holger Rumpold.

  Anti-EGFR targeting is one of the key strategies in the treatment of metastatic colorectal cancer (mCRC). For almost two decades oncologists have struggled to implement EGFR antibodies in the mCRC continuum of care. Both sidedness and RAS mutational status rank high among the predictive factors for the clinical efficacy of EGFR inhibitors. A prospective phase III trial has recently confirmed that anti-EGFR targeting confers an overall survival benefit only in left sided RAS-wildtype tumors when given in first line. It is a matter of discussion if more clinical benefit can be reached by considering putative primary resistance mechanisms (e.g., HER2, BRAF, PIK3CA, etc.) at this early stage of treatment. The value of this procedure in daily routine clinical utility has not yet been clearly delineated. Re-exposure to EGFR antibodies becomes increasingly crucial in the disease journey of mCRC. Yet re- induction or re-challenge strategies have been problematic as they relied on mathematical models that described the timely decay of EGFR antibody resistant clones. The advent of liquid biopsy and the implementation of more accurate next-generation sequencing (NGS) based high throughput methods allows for tracing of EGFR resistant clones in real time. These displays the spatiotemporal heterogeneity of metastatic disease compared to the former standard radiographic assessment and re-biopsy. These techniques may move EGFR inhibition in mCRC into the area of precision medicine in order to apply EGFR antibodies with the increase or decrease of EGFR resistant clones. This review critically discusses established concepts of tackling the EGFR pathway in mCRC and provides insight into the growing field of liquid biopsy guided personalized approaches of EGFR inhibition in mCRC.

Keywords:  Metastatic colorectal cancer; clonal evolution; epidermal growth factor receptor; liquid biopsy; maintenance; molecular oncology; systemic treatment

DOI:  https://doi.org/10.3389/fonc.2022.1048166
Trends Cell Biol. 2022 Dec 03. pii: S0962-8924(22)00254-9. [Epub ahead of print]

Targeting ferroptosis to treat colorectal cancer.

Hong Yan, Ronan Talty, Caroline H Johnson.

  Ferroptosis has emerged as a promising target for colorectal cancer (CRC) treatment. Although disrupting glutathione metabolism is the primary strategy for ferroptosis induction, additional key pathways link ferroptosis to CRC pathogenesis. Here, we discuss arachidonic acid (AA), energy metabolism, AMP-activated protein kinase (AMPK), phosphatidylinositol-3-kinase (PI3K)-protein kinase B (Akt)-mammalian target of rapamycin (mTOR), and Hippo signaling, summarize key findings, and propose new conceptual avenues for CRC treatment.

Keywords:  colorectal cancer; ferroptosis; metabolism; therapy

DOI:  https://doi.org/10.1016/j.tcb.2022.11.003
ESMO Open. 2022 Dec 06. pii: S2059-7029(22)00273-3. [Epub ahead of print]7(6): 100639

A first-in-human study of the anti-LAG-3 antibody favezelimab plus pembrolizumab in previously treated, advanced microsatellite stable colorectal cancer.

E Garralda, A Sukari, N J Lakhani, A Patnaik, Y Lou, S-A Im, T Golan, R Geva, M Wermke, M de Miguel, J Palcza, S Jha, M Chaney, A K Abraham, J Healy, G S Falchook.

   BACKGROUND: Treatment options are limited for participants with microsatellite stable (MSS) metastatic colorectal cancer (mCRC) that progressed after two or more prior therapies. Studies have shown that blockade of both lymphocyte-activation gene 3 (LAG-3) and programmed cell death protein 1 (PD-1) can improve antitumor activity. Here, we evaluate the antitumor activity of the LAG-3 antibody favezelimab alone or in combination with pembrolizumab in participants with MSS mCRC.
PATIENTS AND METHODS: Eligible participants with MSS PD-1/programmed death-ligand 1 (PD-L1) treatment-naive mCRC that progressed on two or more prior therapies received 800 mg favezelimab, 800 mg favezelimab plus 200 mg pembrolizumab, or 800 mg favezelimab/200 mg pembrolizumab co-formulation, every 3 weeks. The primary endpoint was safety, the secondary endpoint was objective response rate (ORR), and exploratory endpoints included duration of response, progression-free survival (PFS), and overall survival (OS).
RESULTS: At the data cut-off date of 23 October 2020, a total of 20 participants received favezelimab alone, 89 received favezelimab plus pembrolizumab (including as favezelimab/pembrolizumab co-formulation); 48 had PD-L1 combined positive score (CPS) ≥1 tumors. At this interim analysis median follow-up was 5.8 months with favezelimab and 6.2 with favezelimab plus pembrolizumab. Treatment-related adverse events (TRAEs) were 65% with favezelimab and 65.2% with favezelimab plus pembrolizumab. Grade ≥3 TRAEs were 15% with favezelimab and 20% with favezelimab plus pembrolizumab. No grade 5 TRAEs occurred. Common TRAEs (≥15%) included fatigue (20.0%), nausea (15.0%) with favezelimab, and fatigue (16.9%) with favezelimab plus pembrolizumab. Confirmed ORR was 6.3% with favezelimab plus pembrolizumab, with median duration of response of 10.6 months (range 5.6-12.7 months), median OS of 8.3 months (95% confidence interval 5.5-12.9 months), and median PFS of 2.1 months (1.9-2.2 months). In an exploratory analysis of PD-L1 CPS ≥1 tumors, the confirmed ORR was 11.1%, median OS was 12.7 months (4.5 to not reached), and median PFS was 2.2 months (1.8-4.2 months) with favezelimab plus pembrolizumab.
CONCLUSIONS: Favezelimab with or without pembrolizumab had a manageable safety profile, with no treatment-related deaths. Promising antitumor activity was observed with combination therapy, particularly in participants with PD-L1 CPS ≥1 tumors.

Keywords:  LAG-3; PD-1; advanced; colorectal cancer

DOI:  https://doi.org/10.1016/j.esmoop.2022.100639
Commun Biol. 2022 Dec 03. 5(1): 1326

An integrated map of fibroblastic populations in human colon mucosa and cancer tissues.

Siying Li, Ran Lu, Linjuan Shu, Yulin Chen, Jin Zhao, Junlong Dai, Qiaorong Huang, Xue Li, Wentong Meng, Feiwu Long, Yuan Li, Chuanwen Fan, Zongguang Zhou, Xianming Mo.

Fibroblasts and myofibroblasts are major mesenchymal cells in the lamina propria of colon mucosa and in colon cancer tissues. Detailed insight into the highly specific populations of fibroblasts and myofibroblasts is required to understand the integrity and homeostasis of human colon mucosa and colon cancer. Based on gene expression profiles of single cells, we identified fibroblast populations that produce extracellular matrix components, Wnt ligand- and BMP-secreting fibroblasts, chemokine- and chemokine ligand-generating fibroblasts, highly activated fibroblasts, immune-modulating fibroblasts, epithelial cell-modulating myofibroblasts, stimuli-responsive myofibroblasts, proliferating myofibroblasts, fibroblast-like myofibroblasts, matrix producing myofibroblasts, and contractile myofibroblasts in human colon mucosa. In colon cancer tissue, the compositions of fibroblasts and myofibroblasts were highly altered, as were the expressing patterns of genes including BMPs, Wnt ligands, chemokines, chemokine ligands, growth factors and extracellular matrix components in fibroblasts and myofibroblasts. Our work expands the working atlas of fibroblasts and myofibroblasts and provides a framework for interrogating the complexity of stromal cells in human healthy colon mucosa and colon cancer tissues.

DOI: https://doi.org/10.1038/s42003-022-04298-5
Front Immunol. 2022 ;13 1032314

Predictive biomarkers of colon cancer immunotherapy: Present and future.

Wanting Hou, Cheng Yi, Hong Zhu.

  Immunotherapy has revolutionized colon cancer treatment. Immune checkpoint inhibitors (ICIs) have shown clinical benefits for colon cancer patients, especially those with high microsatellite instability (MSI-H). In 2020, the US Food and Drug Administration (FDA)-approved ICI pembrolizumab as the first-line treatment for metastatic MSI-H colon cancer patients. Additionally, neoadjuvant immunotherapy has presented efficacy in treating early-stage colon cancer patients. Although MSI has been thought of as an effective predictive biomarker for colon cancer immunotherapy, only a small proportion of colon cancer patients were MSI-H, and certain colon cancer patients with MSI-H presented intrinsic or acquired resistance to immunotherapy. Thus, further search for predictive biomarkers to stratify patients is meaningful in colon cancer immunotherapy. Except for MSI, other biomarkers, such as PD-L1 expression level, tumor mutation burden (TMB), tumor-infiltrating lymphocytes (TILs), certain gut microbiota, ctDNA, and circulating immune cells were also proposed to be correlated with patient survival and ICI efficacy in some colon cancer clinical studies. Moreover, developing new diagnostic techniques helps identify accurate predictive biomarkers for colon cancer immunotherapy. In this review, we outline the reported predictive biomarkers in colon cancer immunotherapy and further discuss the prospects of technological changes for biomarker development in colon cancer immunotherapy.

Keywords:  biomarker; colon cancer; immune checkpoint inhibitor (ICI); immunotherapy; predictive

DOI:  https://doi.org/10.3389/fimmu.2022.1032314
Lancet Gastroenterol Hepatol. 2022 Dec 02. pii: S2468-1253(22)00380-6. [Epub ahead of print]

Low intensity treatment in metastatic colorectal cancer.

Hui-Li Wong, Jeanne Tie.

DOI: https://doi.org/10.1016/S2468-1253(22)00380-6
Am J Physiol Gastrointest Liver Physiol. 2022 Dec 06.

Spatial transcriptomics add a new dimension to our understanding of the gut.

Charles H Danan, Kay Katada, Louis R Parham, Kathryn E Hamilton.

  The profound complexity of the intestinal mucosa demands a spatial approach to the study of gut transcriptomics. Although single-cell RNA sequencing has revolutionized our ability to survey the diverse cell types of the intestine, knowledge of cell type alone cannot fully describe the cells that make up the intestinal mucosa. During homeostasis and disease, dramatic gradients of oxygen, nutrients, extracellular matrix proteins, morphogens, and microbiota collectively dictate intestinal cell state, and only spatial techniques can articulate differences in cellular transcriptomes at this level. Spatial transcriptomic techniques assign transcriptomic data to precise regions in a tissue of interest. In recent years, these protocols have become increasingly accessible, and their application in the intestinal mucosa has exploded in popularity. In the gut, spatial transcriptomics typically involve the application of tissue sections to spatially barcoded RNA sequencing or laser capture microdissection followed by RNA sequencing. In combination with single-cell RNA sequencing, these spatial sequencing approaches allow for the construction of spatial transcriptional maps at pseudo-single-cell resolution. In this review, we describe the spatial transcriptomic technologies recently applied in the gut and the previously unattainable discoveries that they have produced.

Keywords:  RNA; gut; intestine; spatial; transcriptomics

DOI:  https://doi.org/10.1152/ajpgi.00191.2022
Front Immunol. 2022 ;13 1057181

Cytokine-mediated crosstalk between cancer stem cells and their inflammatory niche from the colorectal precancerous adenoma stage to the cancerous stage: Mechanisms and clinical implications.

Guanglin Cui, Ziqi Wang, Hanzhe Liu, Zhigang Pang.

  The majority of colorectal cancers (CRCs) are thought to arise from precancerous adenomas. Upon exposure to diverse microenvironmental factors, precancerous stem cells (pCSCs) undergo complex genetic/molecular changes and gradually progress to form cancer stem cells (CSCs). Accumulative evidence suggests that the pCSC/CSC niche is an inflammatory dominated milieu that contains different cytokines that function as the key communicators between pCSCs/CSCs and their niche and have a decisive role in promoting CRC development, progression, and metastasis. In view of the importance and increasing data about cytokines in modulating pCSCs/CSC stemness properties and their significance in CRC, this review summarizes current new insights of cytokines, such as interleukin (IL)-4, IL-6, IL-8, IL-17A, IL-22, IL-23, IL-33 and interferon (IFN)-γ, involving in the modulation of pCSC/CSC properties and features in precancerous and cancerous lesions and discusses the possible mechanisms of adenoma progression to CRCs and their therapeutic potential.

Keywords:  cancer stem cells; colorectum; cytokine; stem cell; tumorigenesis

DOI:  https://doi.org/10.3389/fimmu.2022.1057181