bims-instec 2022-07-10 papers

Nature. 2022 Jul 07.

Cell-matrix interface regulates dormancy in human colon cancer stem cells.

Yuki Ohta, Masayuki Fujii, Sirirat Takahashi, Ai Takano, Kosaku Nanki, Mami Matano, Hikaru Hanyu, Megumu Saito, Mariko Shimokawa, Shingo Nishikori, Yoshiko Hatano, Ryota Ishii, Kazuaki Sawada, Akihito Machinaga, Wataru Ikeda, Takeshi Imamura, Toshiro Sato.

Cancer relapse after chemotherapy remains a main cause of cancer-related death. Although the relapse is thought to result from the propagation of resident cancer stem cells (CSCs)1, a lack of experimental platforms that enable prospective analysis of CSC dynamics with sufficient spatiotemporal resolution has hindered testing of this hypothesis. Here, we develop a live genetic lineage-tracing system that allows longitudinal tracking of individual cells in xenotransplanted human colorectal cancer organoids and identify LGR5+ CSCs that display a dormant behavior in a chemo-naive state. Dormant LGR5+ cells are marked by p27 expression, and intravital imaging directly demonstrates the persistence of LGR5+p27+ cells during chemotherapy, followed by clonal expansion. Transcriptome analysis reveals an upregulation of COL17A1, a cell adhesion molecule that strengthens hemidesmosome, in dormant LGR5+p27+ cells. COL17A1-knockout organoids lose the dormant LGR5+p27+ subpopulation and become sensitive to chemotherapy, suggesting a role of cell-matrix interface in dormancy maintenance. Chemotherapy disrupts COL17A1 and breaks the dormancy in LGR5+p27+ cells through FAK-YAP activation. Abrogation of YAP signaling restrains chemo-resistant cells from exiting dormancy and delays tumor regrowth, highlighting the therapeutic potential of YAP inhibition in preventing cancer relapse. These results offer a viable therapeutic approach to overcome refractoriness of human colorectal cancer to conventional chemotherapy.

DOI: https://doi.org/10.1038/s41586-022-05043-y

Anticancer Res. 2022 Jul;42(7): 3325-3340

PI3K/Akt and Wnt/β-catenin Signaling Cross-regulate NF-κB Signaling in TNF-α-induced Human Lgr5+ Intestinal Stem Cells.

Xiaopeng Zhao, Binbin Ma, Hongli Zhu, Jun Bai, Lixiu Liu, Xiaoning Li, Jing Cai, Beibei Wang, Lijun Wang, Yuan Pang, Hui Zhang, Y I Yang, Nan Li, Mei Chen.

BACKGROUND/AIM: Intestinal stem cells (ISCs) are responsible for intestinal proliferation, differentiation, and neoplasia, and also play a crucial role in inflammation. Thus, it is important to investigate the effect of TNF-α on the activities of NF-κB, PI3K/Akt, and Wnt/β-catenin signaling pathways.
MATERIALS AND METHODS: The Lgr5+ intestinal cells were isolated using fluorescence-activated cell sorting from NCM460 spheroid cells, and the potential molecular mechanisms were investigated via short hairpin RNA (shRNA) transfection or the use of an inhibitor.
RESULTS: The Lgr5+ cells were termed ISCs because of the higher expression of stem cell genes, including Sox2, Nanog, Oct4, Lgr5, and CD133. The Lgr5+ ISCs had a higher proliferation capacity, invasive ability, and drug resistance to 5-fluorouracil, as well as higher expression levels of anti-apoptotic proteins but lower expression levels of pro-apoptotic proteins, compared with Lgr5~ cells. The PI3K/Akt and Wnt/β-catenin pathways were triggered by the TNF-α-induced activation of NF-κB signaling. Notably, when p65 expression was knocked-down via shRNA transfection in Lgr5+ ISCs, the TNF-α-induced activation of the NF-κB, PI3K/Akt, and Wnt/β-catenin pathways were reversed. The same effect was observed with regards to β-catenin shRNA transfection. Moreover, the Akt inhibitor MK2206 inhibited the TNF-α-induced activation of the PI3K/Akt pathway, as well as the NF-κB and Wnt/β-catenin pathways.
CONCLUSION: PI3K/Akt and Wnt/β-catenin signaling cross-regulate NF-κB signaling in TNF-α-induced human Lgr5+ ISCs.

Keywords: TNF-α; cross-regulation; intestinal stem cells; signaling pathway

DOI: https://doi.org/10.21873/anticanres.15820

Int J Mol Sci. 2022 Jun 24. pii: 7055. [Epub ahead of print]23(13):

Stem Cell Models for Cancer Therapy.

Nitin Telang.

Metastatic progression of female breast and colon cancer represents a major cause of mortality in women. Spontaneous/acquired resistance to conventional and targeted chemo-endocrine therapy is associated with the emergence of drug-resistant tumor-initiating cancer stem cell populations. The cancer-initiating premalignant stem cells exhibit activation of select cancer cell signaling pathways and undergo epithelial-mesenchymal transition, leading to the evolution of a metastatic phenotype. The development of reliable cancer stem cell models provides valuable experimental approaches to identify novel testable therapeutic alternatives for therapy-resistant cancer. Drug-resistant stem cell models for molecular subtypes of clinical breast cancer and for genetically predisposed colon cancer are developed by selecting epithelial cells that survive in the presence of cytostatic concentrations of relevant therapeutic agents. These putative stem cells are characterized by the expression status of select cellular and molecular stem cell markers. The stem cell models are utilized as experimental approaches to examine the stem-cell-targeted growth inhibitory efficacy of naturally occurring dietary phytochemicals. The present review provides a systematic discussion on (i) conceptual and experimental aspects relevant to the chemo-endocrine therapy of breast and colon cancer, (ii) molecular/cellular aspects of cancer stem cells and (iii) potential stem-cell-targeting lead compounds as testable alternatives against the progression of therapy-resistant breast and colon cancer.

Keywords: breast and colon cancer; stem cells; testable therapeutic alternatives

DOI: https://doi.org/10.3390/ijms23137055

Front Immunol. 2022 ;13 887048

Identification of Genes Related to 5-Fluorouracil Based Chemotherapy for Colorectal Cancer.

Xingxing Huang, Kun Ke, Weiwei Jin, Qianru Zhu, Qicong Zhu, Ruyi Mei, Ruonan Zhang, Shuxian Yu, Lan Shou, Xueni Sun, Jiao Feng, Ting Duan, Yiping Mou, Tian Xie, Qibiao Wu, Xinbing Sui.

Background: Colorectal cancer (CRC) is one of the most common malignancies and its incidence and mortality are increasing yearly. 5-Fluorouracil (5-FU) has long been used as a standard first-line treatment for CRC patients. Although 5-FU-based chemotherapy is effective for advanced CRC, the consequent resistance remains a key problem and causes the poor prognosis of CRC patients. Thus, there is an urgent need to identify new biomarkers to predict the response to 5-FU-based chemotherapy.
Methods: CRC samples were retrieved from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). The immune-related genes were retrieved from the ImmPort database. Single-cell sequencing results from colorectal cancer were obtained by the ArrayExpress database. 5-FU resistance-related genes were filtered and validated by R packages. ESTIMATE algorithms were used to assess the tumor microenvironment (TME). KEGG and GO analysis were performed to explore the biological signaling pathway for resistant-response patients and sensitive-response patients in the tumor microenvironment. pRRophetic algorithms were used to predict 5-FU sensitivity. GSEA and GSVA analysis was performed to excavate the biological signaling pathway of the RBP7 gene.
Results: Nine immune-related genes were identified to be associated with 5-FU resistance and poor disease-free survival (DFS) of CRC patients and the signature of these genes was developed in a DFS-prognostic model. Four immune-related genes were determined to be associated with 5-FU resistance and overall survival (OS) of CRC patients. The signature of these genes was developed an OS-prognostic model. ESTIMATE scores showed a significant difference between 5-FU resistant and 5-FU sensitive CRC patients. Resistant-response patients and sensitive-response patients to 5-FU based chemotherapy showed different GO and KEGG enrichment on the tumor microenvironment. RBP7, as a tumor immune microenvironment (TIME) related gene, was found to have the potential of predicting chemotherapy resistance and poor prognosis of CRC patients. GSEA analysis showed multiple signaling differences between the high and low expression of RBP7 in CRC patients. Hypoxia and TNFα signaling via NFκB gene sets were significantly different between chemotherapy resistant (RBP7High) and chemotherapy sensitive (RBP7Low) patients. Single-cell RNA-seq suggested RBP7 was centrally distributed in endothelial stalk cells, endothelial tip cells, and myeloid cells.
Conclusions: Immune-related genes will hopefully be potential prognostic biomarkers to predict chemotherapy resistance for CRC. RBP7 may function as a tumor microenvironment regulator to induce 5-FU resistance, thereby affecting the prognosis of CRC patients.

Keywords: 5-FU resistance; colorectal cancer; immune-related genes; prognosis; tumor microenvironment

DOI: https://doi.org/10.3389/fimmu.2022.887048

Cancers (Basel). 2022 Jun 23. pii: 3081. [Epub ahead of print]14(13):

Intestinal FXR Activation via Transgenic Chimera or Chemical Agonism Prevents Colitis-Associated and Genetically-Induced Colon Cancer.

Marica Cariello, Roberta Zerlotin, Emanuela Pasculli, Elena Piccinin, Claudia Peres, Emanuele Porru, Aldo Roda, Raffaella Maria Gadaleta, Antonio Moschetta.

The Farnesoid X Receptor (FXR) is the master regulator of Bile Acids (BA) homeostasis orchestrating their synthesis, transport and metabolism. Disruption of BA regulation has been linked to gut-liver axis diseases such as colorectal cancer (CRC). In this study, firstly we examined the role of constitutive activation of intestinal FXR in CRC; then we pre-clinically investigated the therapeutic potential of a diet enriched with a synthetic FXR agonist in two models of CRC (chemically-induced and genetic models). We demonstrated that mice with intestinal constitutive FXR activation are protected from AOM/DSS-induced CRC with a significant reduction of tumor number compared to controls. Furthermore, we evaluated the role of chemical FXR agonism in a DSS model of colitis in wild type (WT) and FXRnull mice. WT mice administered with the FXR activating diet showed less morphological alterations and decreased inflammatory infiltrates compared to controls. The FXR activating diet also protected WT mice from AOM/DSS-induced CRC by reducing tumors' number and size. Finally, we proved that the FXR activating diet prevented spontaneous CRC in APCMin/+ mice via an FXR-dependent modulation of BA homeostasis. Our results demonstrate that intestinal FXR activation prevented both inflammation- and genetically-driven colorectal tumorigenesis by modulating BA pool size and composition. This could open new avenues for the therapeutic management of intestinal inflammation and tumorigenesis.

Keywords: Farnesoid X Receptor (FXR); colitis; colon cancer

DOI: https://doi.org/10.3390/cancers14133081

BMC Genom Data. 2022 Jul 06. 23(1): 51

Identification of prognostic stemness biomarkers in colon adenocarcinoma drug resistance.

Ziyue Li, Jierong Chen, Dandan Zhu, Xiaoxiao Wang, Jace Chen, Yu Zhang, Qizhou Lian, Bing Gu.

BACKGROUND: Colon adenocarcinoma (COAD) is one of the leading causes of death worldwide. Cancer stem cells (CSCs) are vital for COAD chemoresistance and recurrence, however little is known about stem cell-related biomarkers in drug resistance and COAD prognosis prediction.
METHODS: To uncover the roles of CSC in COAD tumorigenesis, chemoresistance, and prognosis, we retrieved COAD patients' RNAseq data from TCGA (The Cancer Genome Atlas). We further performed analysis of differentially expressed genes (DEGs) and mRNA expression-based stemness index (mRNAsi) to identify stemness-related COAD biomarkers. We then evaluated the roles of mRNAsi in tumorigenesis, clinical-stage, overall survival (OS), and chemoresistance. Afterward, we used identified prognostic stemness-related genes (PSRGs) to construct a prediction model. After constructing the prediction model, we used elastic Net regression and area under the curve (AUC) to explore the prediction value of PSRGs based on risk scores and the receiver operator characteristic (ROC) curve. To elucidate the underlying interconnected systems, we examined relationships between the levels of TFs, PSRGs, and 50 cancer hallmarks by a Pearson correlation analysis.
RESULTS: Twelve thousand one hundred eight DEGs were identified by comparing 456 primary COADs and 41 normal solid tissue samples. Furthermore, we identified 4351 clinical stage-related DEGs, 16,516 stemness-associated DEGs, and 54 chemoresistance-related DEGs from cancer stages: mRNAsi, and COAD chemoresistance. Compared to normal tissue samples, mRNAsi in COAD patients were marked on an elevation and involved in prognosis (p = 0.027), stemness-related DEGs based on chemoresistance (OR = 3.28, p ≤ 0.001) and AJCC clinical stage relating (OR = 4.02, p ≤ 0.001) to COAD patients. The prediction model of prognosis were constructed using the 6 PSRGs with high accuracy (AUC: 0.659). The model identified universal correlation between NRIP2 and FDFT1 (key PRSGs), and some cancer related transcription factors (TFs) and trademarks of cancer gene were in the regulatory network.
CONCLUSION: We found that mRNAsi is a reliable predictive biomarker of tumorigenesis and COAD prognosis. Our established prediction model of COAD chemoresistance, which includes the six PSRGs, is effective, as the model provides promising therapeutic targets in the COAD.

Keywords: Biomarkers; Cancer stem cell; Chemoresistance; Colon adenocarcinoma; Prognosis

DOI: https://doi.org/10.1186/s12863-022-01063-9