bims-instec Biomed News
on Intestinal stem cells and chemoresistance in colon cancer and intestinal regeneration
Issue of 2023‒07‒02
thirteen papers selected by
Maria-Virginia Giolito
Free University of Brussels
  1. The Histone Demethylase KDM5D Drives Sex Differences in Colorectal Cancer.
  2. Laser Ablation and Intravital Microscopy to Study Intestinal Remodeling.
  3. Tracking the growth fate of single cells and isolating slow-growing cells in human colorectal cancer organoids.
  4. A bioprinted 3D gut model with crypt-villus structures to mimic the intestinal epithelial-stromal microenvironment.
  5. In Vitro and In Vivo Models for Metastatic Intestinal Tumors Using Genotype-Defined Organoids.
  6. Caffeine limits expansion of Apc-deficient clones in the intestine by NOTUM inhibition.
  7. Modeling Intestinal Carcinogenesis Using In Vitro Organoid Cultures.
  8. Kinase-targeted therapy in subsets of colorectal cancer.
  9. Inhibition of the bone morphogenetic protein pathway suppresses tumor growth through downregulation of epidermal growth factor receptor in MEK/ERK-dependent colorectal cancer.
  10. PDE4DIP contributes to colorectal cancer growth and chemoresistance through modulation of the NF1/RAS signaling axis.
  11. An epigenetic signature of advanced colorectal cancer metastasis.
  12. Targeting lysine-specific demethylase 1 (KDM1A/LSD1) impairs colorectal cancer tumorigenesis by affecting cancer cells stemness, motility, and differentiation.
  13. High glucose promotes the progression of colorectal cancer by activating the BMP4 signaling and inhibited by glucagon-like peptide-1 receptor agonist.
  1. Cancer Discov. 2023 Jun 30. OF1
    The Histone Demethylase KDM5D Drives Sex Differences in Colorectal Cancer.
      The Y chromosome histone demethylase gene, KDM5D, drives sex differences in colorectal cancer (CRC).
    DOI:  https://doi.org/10.1158/2159-8290.CD-RW2023-100
  2. J Vis Exp. 2023 06 09.
    Laser Ablation and Intravital Microscopy to Study Intestinal Remodeling.
    Dimitrios Laskaris, Maria Azkanaz, Mijke A de Vreij-Kruidenier, Doreen van Rijswoud-Ram, Hendrik A Messal, Jacco van Rheenen.
      Investigating intestinal recovery in vivo is an exquisite technical challenge. A lack of longitudinal imaging protocols has prevented deeper insights into the cell and tissue scale dynamics that orchestrate intestinal regeneration. Here, we describe an intravital microscopy method that locally induces tissue damage at the single crypt scale and follows the regenerative response of the intestinal epithelium in living mice. Single crypts or larger intestinal fields were ablated by a high-intensity multiphoton infrared laser in a time- and space-controlled manner. Subsequent long-term repetitive intravital imaging enabled the tracking of the damaged areas over time and allowed for the monitoring of crypt dynamics during tissue recovery over a period of multiple weeks. Crypt remodeling events such as crypt fission, fusion, and disappearance were observed in the neighboring tissue upon laser-induced damage. This protocol enables the study of crypt dynamics both in homeostatic and pathophysiological settings, such as aging and tumor initiation.
    DOI:  https://doi.org/10.3791/64756
  3. STAR Protoc. 2023 Jun 28. pii: S2666-1667(23)00362-3. [Epub ahead of print]4(3): 102395
    Tracking the growth fate of single cells and isolating slow-growing cells in human colorectal cancer organoids.
    Roberto Coppo, Jumpei Kondo, Kunishige Onuma, Masahiro Inoue.
      Patient-derived tumor organoids are three-dimensionally cultured cancer cells that enable a suitable platform for studying heterogeneity and plasticity of cancer. We present a protocol for tracking the growth fate of single cells and isolating slow-growing cells in human colorectal cancer organoids. We describe steps for organoid preparation and culturing using the cancer-tissue-originating spheroid method, maintaining cell-cell contact throughout. We then detail a single-cell-derived spheroid-forming and growth assay, confirming single-cell plating, monitoring growth over time, and isolating slow-growing cells. For complete details on the use and execution of this protocol, please refer to Coppo et al.1.
    Keywords:  Cancer; Cell Culture; Cell Isolation; Single Cell
    DOI:  https://doi.org/10.1016/j.xpro.2023.102395
  4. Biomater Adv. 2023 Jun 22. pii: S2772-9508(23)00257-1. [Epub ahead of print]153 213534
    A bioprinted 3D gut model with crypt-villus structures to mimic the intestinal epithelial-stromal microenvironment.
    Núria Torras, Jon Zabalo, Eduardo Abril, Albane Carré, María García-Díaz, Elena Martínez.
      The intestine is a complex tissue with a characteristic three-dimensional (3D) crypt-villus architecture, which plays a key role in the intestinal function. This function is also regulated by the intestinal stroma that actively supports the intestinal epithelium, maintaining the homeostasis of the tissue. Efforts to account for the 3D complex structure of the intestinal tissue have been focused mainly in mimicking the epithelial barrier, while solutions to include the stromal compartment are scarce and unpractical to be used in routine experiments. Here we demonstrate that by employing an optimized bioink formulation and the suitable printing parameters it is possible to produce fibroblast-laden crypt-villus structures by means of digital light projection stereolithography (DLP-SLA). This process provides excellent cell viability, accurate spatial resolution, and high printing throughput, resulting in a robust biofabrication approach that yields functional gut mucosa tissues compatible with conventional testing techniques.
    Keywords:  3D bioprinting; 3D microstructure; Epithelial-stromal interactions; GelMA-PEGDA soft hydrogels; Intestinal mucosa model
    DOI:  https://doi.org/10.1016/j.bioadv.2023.213534
  5. Methods Mol Biol. 2023 ;2691 19-30
    In Vitro and In Vivo Models for Metastatic Intestinal Tumors Using Genotype-Defined Organoids.
    Atsuya Morita, Mizuho Nakayama, Hiroko Oshima, Masanobu Oshima.
      It has been established that the accumulation of driver gene mutations causes malignant progression of colorectal cancer (CRC) through positive selection and clonal expansion, similar to Darwin's evolution. Following this multistep tumorigenesis concept, we previously showed the specific mutation patterns for each process of malignant progression, including submucosal invasion, epithelial mesenchymal transition (EMT), intravasation, and metastasis, using genetically engineered mouse and organoid models. However, we also found that certain populations of cancer-derived organoid cells lost malignant characteristics of metastatic ability, although driver mutations were not impaired, and such subpopulations were eliminated from the tumor tissues by negative selection. These organoid model studies have contributed to our understanding of the cancer evolution mechanism. We herein report the in vitro and in vivo experimental protocols to investigate the survival, growth, and metastatic ability of intestinal tumor-derived organoids. The model system will be useful for basic research as well as the development of clinical strategies.
    Keywords:  Collagen gel; Colon cancer; Imaging; Metastasis; Organoids
    DOI:  https://doi.org/10.1007/978-1-0716-3331-1_2
  6. Cell Mol Gastroenterol Hepatol. 2023 Jun 24. pii: S2352-345X(23)00123-6. [Epub ahead of print]
    Caffeine limits expansion of Apc-deficient clones in the intestine by NOTUM inhibition.
    Milou S van Driel, Jasmijn D G Linssen, Dustin J Flanagan, Nikola Vlahov, Lisanne E Nijman, Nina E de Groot, Clara C Elbers, Jan Koster, Owen J Sansom, Louis Vermeulen, Sanne M van Neerven.
      
    DOI:  https://doi.org/10.1016/j.jcmgh.2023.06.008
  7. Methods Mol Biol. 2023 ;2691 55-69
    Modeling Intestinal Carcinogenesis Using In Vitro Organoid Cultures.
    Wing Hei Chan, Diana Micati, Rebekah M Engel, Genevieve Kerr, Reyhan Akhtar, Thierry Jardé, Helen E Abud.
      Mouse models of intestinal carcinogenesis are very powerful tools for studying the impact of specific mutations on tumor initiation and progression. Mutations can be studied both singularly and in combination using conditional alleles that can be induced in a temporal manner. The steps in intestinal carcinogenesis are complex and can be challenging to image in live animals at a cellular level. The ability to culture intestinal epithelial tissue in three-dimensional organoids in vitro provides an accessible system that can be genetically manipulated and easily visualized to assess specific biological impacts in living tissue. Here, we describe methodology for conditional mutation of genes in organoids from genetically modified mice via induction of Cre recombinase induced by tamoxifen or by transient exposure to TAT-Cre protein and subsequent phenotyping of the organoids. This methodology provides a rapid platform for assessing the cellular changes induced by specific mutations in intestinal tissue.
    Keywords:  Conditional mutation; Cre recombinase; Intestinal organoids
    DOI:  https://doi.org/10.1007/978-1-0716-3331-1_5
  8. Oncoscience. 2023 ;10 24-26
    Kinase-targeted therapy in subsets of colorectal cancer.
    Patricia M Gomez Barila, Jan Paul Medema.
      
    Keywords:  colorectal cancer; drug resistance; kinase inhibitors; signaling pathways
    DOI:  https://doi.org/10.18632/oncoscience.580
  9. Cancer Sci. 2023 Jun 25.
    Inhibition of the bone morphogenetic protein pathway suppresses tumor growth through downregulation of epidermal growth factor receptor in MEK/ERK-dependent colorectal cancer.
    Shota Shimizu, Jumpei Kondo, Kunishige Onuma, Roberto Coppo, Kasumi Ota, Mayumi Kamada, Yohei Harada, Yoshihisa Tanaka, Mai Adachi Nakazawa, Yoshinori Tamada, Yasushi Okuno, Kenji Kawada, Kazutaka Obama, Robert J Coffey, Yoshiyuki Fujiwara, Masahiro Inoue.
      The bone morphogenetic protein (BMP) pathway promotes differentiation and induces apoptosis in normal colorectal epithelial cells. However, its role in colorectal cancer (CRC) is controversial, where it can act as context-dependent tumor promoter or tumor suppressor. Here we have found that CRC cells reside in a BMP-rich environment based on curation of two publicly available RNA-sequencing databases. Suppression of BMP using a specific BMP inhibitor, LDN193189, suppresses the growth of select CRC organoids. Colorectal cancer organoids treated with LDN193189 showed a decrease in epidermal growth factor receptor, which was mediated by protein degradation induced by leucine-rich repeats and immunoglobulin-like domains protein 1 (LRIG1) expression. Among 18 molecularly characterized CRC organoids, suppression of growth by BMP inhibition correlated with induction of LRIG1 gene expression. Notably, knockdown of LRIG1 in organoids diminished the growth-suppressive effect of LDN193189. Furthermore, in CRC organoids, which are susceptible to growth suppression by LDN193189, simultaneous treatment with LDN193189 and trametinib, an FDA-approved MEK inhibitor, resulted in cooperative growth inhibition both in vitro and in vivo. Taken together, the simultaneous inhibition of BMP and MEK could be a novel treatment option in CRC cases, and evaluating in vitro growth suppression and LRIG1 induction by BMP inhibition using patient-derived organoids could offer functional biomarkers for predicting potential responders to this regimen.
    Keywords:  BMP; EGFR; LRIG1; colorectal cancer; organoid
    DOI:  https://doi.org/10.1111/cas.15882
  10. Cell Death Dis. 2023 Jun 24. 14(6): 373
    PDE4DIP contributes to colorectal cancer growth and chemoresistance through modulation of the NF1/RAS signaling axis.
    Rulu Pan, Juji Dai, Weicheng Liang, Hongxiao Wang, Lin Ye, Siqi Ye, Ziqi Lin, Shishun Huang, Yan Xiong, Li Zhang, Liting Lu, Ouchen Wang, Xian Shen, Wanqin Liao, Xincheng Lu.
      Phosphodiesterase 4D interacting protein (PDE4DIP) is a centrosome/Golgi protein associated with cyclic nucleotide phosphodiesterases. PDE4DIP is commonly mutated in human cancers, and its alteration in mice leads to a predisposition to intestinal cancer. However, the biological function of PDE4DIP in human cancer remains obscure. Here, we report for the first time the oncogenic role of PDE4DIP in colorectal cancer (CRC) growth and adaptive MEK inhibitor (MEKi) resistance. We show that the expression of PDE4DIP is upregulated in CRC tissues and associated with the clinical characteristics and poor prognosis of CRC patients. Knockdown of PDE4DIP impairs the growth of KRAS-mutant CRC cells by inhibiting the core RAS signaling pathway. PDE4DIP plays an essential role in the full activation of oncogenic RAS/ERK signaling by suppressing the expression of the RAS GTPase-activating protein (RasGAP) neurofibromin (NF1). Mechanistically, PDE4DIP promotes the recruitment of PLCγ/PKCε to the Golgi apparatus, leading to constitutive activation of PKCε, which triggers the degradation of NF1. Upregulation of PDE4DIP results in adaptive MEKi resistance in KRAS-mutant CRC by reactivating the RAS/ERK pathway. Our work reveals a novel functional link between PDE4DIP and NF1/RAS signal transduction and suggests that targeting PDE4DIP is a promising therapeutic strategy for KRAS-mutant CRC.
    DOI:  https://doi.org/10.1038/s41419-023-05885-y
  11. iScience. 2023 Jun 16. 26(6): 106986
    An epigenetic signature of advanced colorectal cancer metastasis.
    Euan J Rodger, Gregory Gimenez, Priyadarshana Ajithkumar, Peter A Stockwell, Suzan Almomani, Sarah A Bowden, Anna L Leichter, Antonio Ahn, Sharon Pattison, John L McCall, Sebastian Schmeier, Frank A Frizelle, Michael R Eccles, Rachel V Purcell, Aniruddha Chatterjee.
      Colorectal cancer (CRC) is a leading cause of morbidity and mortality worldwide. The majority of CRC deaths are caused by tumor metastasis, even following treatment. There is strong evidence for epigenetic changes, such as DNA methylation, accompanying CRC metastasis and poorer patient survival. Earlier detection and a better understanding of molecular drivers for CRC metastasis are of critical clinical importance. Here, we identify a signature of advanced CRC metastasis by performing whole genome-scale DNA methylation and full transcriptome analyses of paired primary cancers and liver metastases from CRC patients. We observed striking methylation differences between primary and metastatic pairs. A subset of loci showed coordinated methylation-expression changes, suggesting these are potentially epigenetic drivers that control the expression of critical genes in the metastatic cascade. The identification of CRC epigenomic markers of metastasis has the potential to enable better outcome prediction and lead to the discovery of new therapeutic targets.
    Keywords:  Cancer; Clinical genetics; DNA methylation; Transcriptomics; epigenetics; epigenomics
    DOI:  https://doi.org/10.1016/j.isci.2023.106986
  12. Cell Death Discov. 2023 Jun 29. 9(1): 201
    Targeting lysine-specific demethylase 1 (KDM1A/LSD1) impairs colorectal cancer tumorigenesis by affecting cancer cells stemness, motility, and differentiation.
    Annamaria Antona, Giovanni Leo, Francesco Favero, Marco Varalda, Jacopo Venetucci, Stefania Faletti, Matilde Todaro, Eleonora Mazzucco, Enrica Soligo, Chiara Saglietti, Giorgio Stassi, Marcello Manfredi, Giuliana Pelicci, Davide Corà, Guido Valente, Daniela Capello.
      Among all cancers, colorectal cancer (CRC) is the 3rd most common and the 2nd leading cause of death worldwide. New therapeutic strategies are required to target cancer stem cells (CSCs), a subset of tumor cells highly resistant to present-day therapy and responsible for tumor relapse. CSCs display dynamic genetic and epigenetic alterations that allow quick adaptations to perturbations. Lysine-specific histone demethylase 1A (KDM1A also known as LSD1), a FAD-dependent H3K4me1/2 and H3K9me1/2 demethylase, was found to be upregulated in several tumors and associated with a poor prognosis due to its ability to maintain CSCs staminal features. Here, we explored the potential role of KDM1A targeting in CRC by characterizing the effect of KDM1A silencing in differentiated and CRC stem cells (CRC-SCs). In CRC samples, KDM1A overexpression was associated with a worse prognosis, confirming its role as an independent negative prognostic factor of CRC. Consistently, biological assays such as methylcellulose colony formation, invasion, and migration assays demonstrated a significantly decreased self-renewal potential, as well as migration and invasion potential upon KDM1A silencing. Our untargeted multi-omics approach (transcriptomic and proteomic) revealed the association of KDM1A silencing with CRC-SCs cytoskeletal and metabolism remodeling towards a differentiated phenotype, supporting the role of KDM1A in CRC cells stemness maintenance. Also, KDM1A silencing resulted in up-regulation of miR-506-3p, previously reported to play a tumor-suppressive role in CRC. Lastly, loss of KDM1A markedly reduced 53BP1 DNA repair foci, implying the involvement of KDM1A in the DNA damage response. Overall, our results indicate that KDM1A impacts CRC progression in several non-overlapping ways, and therefore it represents a promising epigenetic target to prevent tumor relapse.
    DOI:  https://doi.org/10.1038/s41420-023-01502-1
  13. BMC Cancer. 2023 Jun 27. 23(1): 594
    High glucose promotes the progression of colorectal cancer by activating the BMP4 signaling and inhibited by glucagon-like peptide-1 receptor agonist.
    Bingwei Ma, Xingchun Wang, Hui Ren, Yingying Li, Haijiao Zhang, Muqing Yang, Jiyu Li.
      BACKGROUND: The detailed molecular mechanism between type 2 diabetes mellitus (T2DM) and colorectal cancer (CRC) is still uncertain. Bone morphogenetic protein 4 (BMP4) dysregulation is implicated in T2DM and CRC, respectively. This study aims to investigate whether BMP4 can mediate the interaction of CRC with T2DM.METHODS: We firstly explored the expression of BMP4 in The Cancer Genome Altas (TCGA) databases and CRC patients with or without DM from the Shanghai Tenth People's Hospital. The diabetic model of CRC cell lines in vitro and the mice model in vivo were developed to explore the BMP4 expression during CRC with or without diabetes. Further inhibition of BMP4 to observe its effects on CRC. Also, glucagon-like peptide-1 receptor agonist (GLP-1RA) was used to verify the underlying mechanism of hypoglycemic drugs on CRC via BMP4.
    RESULTS: BMP4 expression was upregulated in CRC patients, and significantly higher in CRC patients with diabetes (P < 0.05). High glucose-induced insulin resistance (IR)-CRC cells and diabetic mice with metastasis model of CRC had increased BMP4 expression, activated BMP4-Smad1/5/8 pathway, and improved proliferative and metastatic ability mediated by epithelial-mesenchymal transition (EMT). And, treated CRC cells with exogenously BMP inhibitor-Noggin or transfected with lentivirus (sh-BMP4) could block the upregulated metastatic ability of CRC cells induced by IR. Meanwhile, GLP-1R was downregulated by high glucose-induced IR while unregulated by BMP4 inhibitor noggin, and treated GLP-1RA could suppress the proliferation of CRC cells induced by IR through downregulated BMP4.
    CONCLUSIONS: BMP4 increased by high glucose promoted the EMT of CRC. The mechanism of the BMP4/Smad pathway was related to the susceptible metastasis of high glucose-induced IR-CRC. The commonly used hypoglycemic drug, GLP-1RA, inhibited the growth and promoted the apoptosis of CRC through the downregulation of BMP4. The result of our study suggested that BMP4 might serve as a therapeutic target in CRC patients with diabetes.
    Keywords:  BMP4; Colorectal cancer; EMT; GLP-1RA; T2DM
    DOI:  https://doi.org/10.1186/s12885-023-11077-w
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