bims-instec 2024-08-18 papers

bims-instec

Biomed News

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

Issue of 2024–08–18
23 papers selected by
Maria-Virginia Giolito, Université Catholique de Louvain

Loss of ARID3A perturbs intestinal epithelial proliferation-differentiation ratio and regeneration.
An In Vivo Metastasis Model Using Genotype-Defined Tumor Organoids.
Advancements in Research and Treatment Applications of Patient-Derived Tumor Organoids in Colorectal Cancer.
Neuro-mesenchymal interaction mediated by a β2 adrenergic-nerve growth factor feedforward loop promotes colorectal cancer progression.
Integrative single-cell analysis of human colorectal cancer reveals patient stratification with distinct immune evasion mechanisms.
Probing the killing potency of tumor-infiltrating lymphocytes on microarrayed colorectal cancer tumoroids.
A Novel CRISPR/Cas9-mediated mouse model of colon carcinogenesis.
Oncofetal morphogenesis similar to embryonic gut formation by a subpopulation of DLD-1 human colon cancer cells.
5-Fluorouracil resistance-based immune-related gene signature for COAD prognosis.
Human organoids with an autologous tissue-resident immune compartment.
Optimized Protocol for Intestinal Swiss Rolls and Immunofluorescent Staining of Paraffin Embedded Tissue.
The ratio of PKM1/PKM2 is the key factor affecting the glucose metabolism and biological function of colorectal cancer cells.
CCL19-producing fibroblasts promote tertiary lymphoid structure formation enhancing anti-tumor IgG response in colorectal cancer liver metastasis.
Palmitoyltransferase ZDHHC6 promotes colon tumorigenesis by targeting PPARγ-driven lipid biosynthesis via regulating lipidome metabolic reprogramming.
Adapt and shape: metabolic features within the metastatic niche.
Interferon-gamma signaling drives epithelial TNF-alpha receptor-2 expression during colonic tissue repair.
Dysfunctional tumor-infiltrating Vδ1 + T lymphocytes in microsatellite-stable colorectal cancer.
CFTR and colorectal cancer susceptibility: an urgent need for further studies.
The Role of Adipocytes Recruited as Part of Tumor Microenvironment in Promoting Colorectal Cancer Metastases.
SLC12A8 upregulation promotes colorectal cancer progression and chemoresistance.
RPL22 is a tumor suppressor in MSI-high cancers and a splicing regulator of MDM4.
Integrative plasma and fecal metabolomics identify functional metabolites in adenoma-colorectal cancer progression and as early diagnostic biomarkers.
The role of cancer-associated fibroblasts in the invasion and metastasis of colorectal cancer.

J Exp Med. 2024 Oct 07. pii: e20232279. [Epub ahead of print]221(10):

Loss of ARID3A perturbs intestinal epithelial proliferation-differentiation ratio and regeneration.

Nikolaos Angelis, Anna Baulies, Florian Hubl, Anna Kucharska, Gavin Kelly, Miriam Llorian, Stefan Boeing, Vivian S W Li.

Intestinal stem cells at the crypt divide and give rise to progenitor cells that proliferate and differentiate into various mature cell types in the transit-amplifying (TA) zone. Here, we showed that the transcription factor ARID3A regulates intestinal epithelial cell proliferation and differentiation at the TA progenitors. ARID3A forms an expression gradient from the villus tip to the upper crypt mediated by TGF-β and WNT. Intestinal-specific deletion of Arid3a reduces crypt proliferation, predominantly in TA cells. Bulk and single-cell transcriptomic analysis shows increased enterocyte and reduced secretory differentiation in the Arid3a cKO intestine, accompanied by enriched upper-villus gene signatures of both cell lineages. We find that the enhanced epithelial differentiation in the Arid3a-deficient intestine is caused by increased binding and transcription of HNF1 and HNF4. Finally, we show that loss of Arid3a impairs irradiation-induced regeneration with sustained cell death and reprogramming. Our findings imply that Arid3a functions to fine-tune the proliferation-differentiation dynamics at the TA progenitors, which are essential for injury-induced regeneration.

DOI: https://doi.org/10.1084/jem.20232279
Methods Mol Biol. 2024 ;2828 57-68

An In Vivo Metastasis Model Using Genotype-Defined Tumor Organoids.

Atsuya Morita, Mizuho Nakayama, Hiroko Oshima, Masanobu Oshima.

  Recent cancer genome analyses have identified frequently mutated genes that are responsible for the development and malignant progression of cancers, including colorectal cancer (CRC). We previously constructed mouse models that carried major driver mutations of CRC, namely Apc, Kras, Tgfbr2, Trp53, and Fbxw7, in combinations. Comprehensive histological analyses of the models showed a link between mutation combinations and malignant phenotypes, such as invasion, epithelial-mesenchymal transition (EMT), and metastasis. The major cause of cancer-related death is metastasis, making it important to understand the mechanism underlying metastasis in order to develop novel therapeutic strategies. To this end, we have established intestinal tumor-derived organoids from different genotyped mice and generated liver metastasis models via transplantation of the organoids into the spleen. Through histological and imaging analyses of the transplantation models, we have determined that the combination of Apc, Kras, Tgfbr2, and Trp53 mutations promotes liver metastasis at a high incidence. We also demonstrated polyclonal metastasis of tumor cell clusters consisting of genetically and phenotypically distinct cells through our model analysis. These organoid transplantation models recapitulate human CRC metastasis, constituting a useful tool for basic and clinical cancer research as a preclinical model. We herein report the experimental protocols of the organoid culture and generation of metastasis models.

Keywords:  Colon cancer; Imaging; Liver metastasis; Mouse models; Organoids

DOI:  https://doi.org/10.1007/978-1-0716-4023-4_6
Cancers (Basel). 2024 Jul 26. pii: 2671. [Epub ahead of print]16(15):

Advancements in Research and Treatment Applications of Patient-Derived Tumor Organoids in Colorectal Cancer.

Denise van der Graaff, Sofie Seghers, Pieterjan Vanclooster, Christophe Deben, Timon Vandamme, Hans Prenen.

  Colorectal cancer (CRC) remains a significant health burden globally, being the second leading cause of cancer-related mortality. Despite significant therapeutic advancements, resistance to systemic antineoplastic agents remains an important obstacle, highlighting the need for innovative screening tools to tailor patient-specific treatment. This review explores the application of patient-derived tumor organoids (PDTOs), three-dimensional, self-organizing models derived from patient tumor samples, as screening tools for drug resistance in CRC. PDTOs offer unique advantages over traditional models by recapitulating the tumor architecture, cellular heterogeneity, and genomic landscape and are a valuable ex vivo predictive drug screening tool. This review provides an overview of the current literature surrounding the use of PDTOs as an instrument for predicting therapy responses in CRC. We also explore more complex models, such as co-cultures with important stromal cells, such as cancer-associated fibroblasts, and organ-on-a-chip models. Furthermore, we discuss the use of PDTOs for drug repurposing, offering a new approach to identify the existing drugs effective against drug-resistant CRC. Additionally, we explore how PDTOs serve as models to gain insights into drug resistance mechanisms, using newer techniques, such as single-cell RNA sequencing and CRISPR-Cas9 genome editing. Through this review, we aim to highlight the potential of PDTOs in advancing our understanding of predicting therapy responses, drug resistance, and biomarker identification in CRC management.

Keywords:  cancer; colorectal cancer; drug resistance; organoids; patient-derived tumor organoids; precision medicine

DOI:  https://doi.org/10.3390/cancers16152671
Cancer Discov. 2024 Aug 13.

Neuro-mesenchymal interaction mediated by a β2 adrenergic-nerve growth factor feedforward loop promotes colorectal cancer progression.

Hiroki Kobayashi, Tadashi Iida, Yosuke Ochiai, Ermanno Malagola, Xiaofei Zhi, Ruth A White, Jin Qian, Feijing Wu, Quin T Waterbury, Ruhong Tu, Biyun Zheng, Jonathan S LaBella, Leah B Zamechek, Atsushi Ogura, Susan L Woods, Daniel L Worthley, Atsushi Enomoto, Timothy C Wang.

Cancer-associated fibroblasts (CAFs) and nerves, components of the tumor microenvironment, have each been shown to directly promote gastrointestinal cancers. However, it remains unknown whether these cells interact with each other to regulate cancer progression. We found that in colorectal cancer (CRC) norepinephrine induces ADRB2-dependent nerve growth factor (NGF) secretion from CAFs, which in turn increases intra-tumor sympathetic innervation and norepinephrine accumulation. Adrenergic stimulation accelerates CRC growth through ADRA2A/Gi-mediated activation of Yes-Associated Protein (YAP). NGF from CAFs directly enhances CRC cell growth via the PI3K/AKT pathway. Treatment with a tropomyosin receptor kinase (Trk) inhibitor decreased YAP and AKT activation and CRC progression in mice. In human CRC, high NGF expression is associated with the mesenchymal-like tumor subtype and poor patient survival. These findings suggest a central role for reciprocal CAF-nerve crosstalk in promoting CRC progression. Blocking this feedforward loop with a Trk inhibitor may represent a potential therapeutic approach for CRC.

DOI: https://doi.org/10.1158/2159-8290.CD-24-0287
Nat Cancer. 2024 Aug 15.

Integrative single-cell analysis of human colorectal cancer reveals patient stratification with distinct immune evasion mechanisms.

Xiaojing Chu, Xiangjie Li, Yu Zhang, Guohui Dang, Yuhui Miao, Wenbin Xu, Jinyu Wang, Zemin Zhang, Sijin Cheng.

The tumor microenvironment (TME) considerably influences colorectal cancer (CRC) progression, therapeutic response and clinical outcome, but studies of interindividual heterogeneities of the TME in CRC are lacking. Here, by integrating human colorectal single-cell transcriptomic data from approximately 200 donors, we comprehensively characterized transcriptional remodeling in the TME compared to noncancer tissues and identified a rare tumor-specific subset of endothelial cells with T cell recruitment potential. The large sample size enabled us to stratify patients based on their TME heterogeneity, revealing divergent TME subtypes in which cancer cells exploit different immune evasion mechanisms. Additionally, by associating single-cell transcriptional profiling with risk genes identified by genome-wide association studies, we determined that stromal cells are major effector cell types in CRC genetic susceptibility. In summary, our results provide valuable insights into CRC pathogenesis and might help with the development of personalized immune therapies.

DOI: https://doi.org/10.1038/s43018-024-00807-z
NPJ Precis Oncol. 2024 Aug 14. 8(1): 179

Probing the killing potency of tumor-infiltrating lymphocytes on microarrayed colorectal cancer tumoroids.

Devanjali Dutta, L Francisco Lorenzo-Martín, François Rivest, Nicolas Broguiere, Lucie Tillard, Simone Ragusa, Nathalie Brandenberg, Sylke Höhnel, Damien Saugy, Sylvie Rusakiewicz, Krisztian Homicsko, George Coukos, Matthias P Lutolf.

Immunotherapy has emerged as a new standard of care for certain cancer patients with specific cellular and molecular makeups. However, there is still an unmet need for ex vivo models able to readily assess the effectiveness of immunotherapeutic treatments in a high-throughput and patient-specific manner. To address this issue, we have developed a microarrayed system of patient-derived tumoroids with recreated immune microenvironments that are optimized for the high-content evaluation of tumor-infiltrating lymphocyte functionality. Here we show that this system offers unprecedented opportunities to evaluate tumor immunogenicity, characterize the response to immunomodulators, and explore novel approaches for personalized immuno-oncology.

DOI: https://doi.org/10.1038/s41698-024-00661-3
Cell Mol Gastroenterol Hepatol. 2024 Aug 09. pii: S2352-345X(24)00145-0. [Epub ahead of print] 101390

A Novel CRISPR/Cas9-mediated mouse model of colon carcinogenesis.

Hajime Kashima, Anthony Fischer, Daniel A Veronese-Paniagua, Vered A Gazit, Changqing Ma, Yan Yan, Marc S Levin, Blair B Madison, Deborah C Rubin.

   BACKGROUND & AIMS: Human sporadic colorectal cancer (CRC) results from a multistep pathway with sequential acquisition of specific genetic mutations in the colorectal epithelium. Modeling CRC in vivo is critical for understanding the tumor microenvironment. To accurately recapitulate human CRC pathogenesis, mouse models must include these multi-step genetic abnormalities.
AIMS: Generate a sporadic CRC model that more closely mimics this multi-step process and use this model to study the role of a novel Let7 target PLAGL2 in CRC pathogenesis.
METHODS: We generated a CRISPR/Cas9 somatic mutagenesis mouse model that is inducible and multiplexed for simultaneous inactivation of multiple genes involved in CRC pathogenesis. We used both a doxycycline-inducible transcriptional activator and a dox-inactivated transcriptional repressor to achieve tight, non-leaky expression of the Cas9 nickase. This mouse has transgenic expression of multiple guide RNAs to induce sporadic inactivation in the gut epithelium of four tumor suppressor genes commonly mutated in CRC, Apc, Pten, Smad4 and Trp53. These were crossed to Vil-LCL-PLAGL2 mice which have Cre-inducible overexpression of PLAGL2 in the gut epithelium.
RESULTS: These mice exhibited random somatic mutations in all four targeted tumor suppressor genes, resulting in multiple adenomas and adenocarcinomas in the small bowel and colon. Crosses with Vil-LCL-PLAGL2 mice demonstrated that gut-specific PLAGL2 overexpression increased colon tumor growth.
CONCLUSIONS: This conditional model represents a new CRISPR/Cas9-mediated mouse model of colorectal carcinogenesis. These mice can be used to investigate the role of novel, previously uncharacterized genes in CRC, in the context of multiple commonly mutated tumor suppressor genes and thus more closely mimic human CRC pathogenesis.

Keywords:  CRISPR/Cas9; PLAGL2; colorectal neoplasia; mouse models

DOI:  https://doi.org/10.1016/j.jcmgh.2024.101390
Exp Cell Res. 2024 Aug 09. pii: S0014-4827(24)00279-9. [Epub ahead of print] 114188

Oncofetal morphogenesis similar to embryonic gut formation by a subpopulation of DLD-1 human colon cancer cells.

Kaoru Miyazaki, Daisuke Hoshino, Rika Kasajima, Shiro Koizume, Naohiko Koshikawa, Yohei Miyagi.

  Cancer stem cells (CSC) are thought to be responsible for cancer phenotypes and cellular heterogeneity. Here we demonstrate that the human colon cancer cell line DLD1 contains two types of CSC-like cells that undergo distinct morphogenesis in the reconstituted basement membrane gel Matrigel. In our method with cancer cell spheroids, the parent cell line (DLD1-P) developed grape-like budding structures, whereas the other (DLD1-Wm) and its single-cell clones dynamically developed worm-like ones. Gene expression analysis suggested that the former mimicked intestinal crypt-villus morphogenesis, while the latter mimicked embryonic hindgut development. The organoids of DLD1-Wm cells rapidly extended in two opposite directions by expressing dipolar proteolytic activity. The invasive morphogenesis required the expression of MMP-2 and CD133 genes and ROCK activity. These cells also exhibited gastrula-like morphogenesis even in two-dimensional cultures without Matrigel. Moreover, the two DLD1 cell lines showed clear differences in cellular growth, tumor growth and susceptibility to paclitaxel. This study also provides a simple organoid culture method for human cancer cell lines. HT-29 and other cancer cell lines underwent characteristic morphogenesis in direct contact with normal fibroblasts. Such organoid cultures would be useful for investigating the nature of CSCs and for screening anti-cancer drugs. Our results lead to the hypothesis that CSC-like cells with both invasive activity and a fetal phenotype, i.e,. oncofetal CSCs, are generated in some types of colon cancers.

Keywords:  Matrigel; cancer stem cells; colon cancer; morphogenesis; oncofetal phenotypes

DOI:  https://doi.org/10.1016/j.yexcr.2024.114188
Heliyon. 2024 Jul 30. 10(14): e34535

5-Fluorouracil resistance-based immune-related gene signature for COAD prognosis.

Haixia Yan, Qinling Ou, Yonglong Chang, Jinhui Liu, Linzi Chen, Duanyang Guo, Sifang Zhang.

   Background: Drug resistance is the primary obstacle to advanced tumor therapy and the key risk factor for tumor recurrence and death. 5-Fluorouracil (5-FU) chemotherapy is the most common chemotherapy for individuals with colorectal cancer, despite numerous options.
Methods: The Gene Expression Omnibus database was utilized to extract expression profile data of HCT-8 human colorectal cancer wild-type cells and their 5-FU-induced drug resistance cell line. These data were used to identify 5-FU resistance-related differentially expressed genes (5FRRDEGs), which intersected with the colorectal adenocarcinoma (COAD) transcriptome data provided by the Cancer Genome Atlas Program database. A prognostic signature containing five 5FRRDEGs (GOLGA8A, KLC3, TIGD1, NBPF1, and SERPINE1) was established after conducting a Cox regression analysis. We conducted nomogram development, drug sensitivity analysis, tumor immune microenvironment analysis, and mutation analysis to assess the therapeutic value of the prognostic qualities.
Results: We identified 166 5FRRDEGs in patients with COAD. Subsequently, we created a prognostic model consisting of five 5FRRDEGs using Cox regression analysis. The patients with COAD were divided into different risk groups by risk score; the high-risk group demonstrated a worse prognosis than the low-risk group.
Conclusion: In summary, the 5FRRDEG-based prognostic model is an effective tool for targeted therapy and chemotherapy in patients with COAD. It can accurately predict the survival prognosis of these patients as well as to provide the direction for exploring the resistance mechanism underlying COAD.

Keywords:  5-FU-Related genes; 5FRRDEGs; Colon adenocarcinoma; Gene signature; Immune infiltration; Prognostic model

DOI:  https://doi.org/10.1016/j.heliyon.2024.e34535
Nature. 2024 Aug 14.

Human organoids with an autologous tissue-resident immune compartment.

Timothy Recaldin, Linda Steinacher, Bruno Gjeta, Marius F Harter, Lukas Adam, Kristina Kromer, Marisa Pimentel Mendes, Marina Bellavista, Mikhail Nikolaev, Giacomo Lazzaroni, Rok Krese, Umut Kilik, Doris Popovic, Bilgenaz Stoll, Régine Gerard, Michael Bscheider, Marc Bickle, Lauriane Cabon, J Gray Camp, Nikolche Gjorevski.

The intimate relationship between the epithelium and immune system is crucial for maintaining tissue homeostasis, with perturbations therein linked to autoimmune disease and cancer1-3. Whereas stem cell-derived organoids are powerful models of epithelial function4, they lack tissue-resident immune cells that are essential for capturing organ-level processes. We describe human intestinal immuno-organoids (IIOs), formed through self-organization of epithelial organoids and autologous tissue-resident memory T (TRM) cells, a portion of which integrate within the epithelium and continuously survey the barrier. TRM cell migration and interaction with epithelial cells was orchestrated by TRM cell-enriched transcriptomic programs governing cell motility and adhesion. We combined IIOs and single-cell transcriptomics to investigate intestinal inflammation triggered by cancer-targeting biologics in patients. Inflammation was associated with the emergence of an activated population of CD8+ T cells that progressively acquired intraepithelial and cytotoxic features. The appearance of this effector population was preceded and potentiated by a T helper-1-like CD4+ population, which initially produced cytokines and subsequently became cytotoxic itself. As a system amenable to direct perturbation, IIOs allowed us to identify the Rho pathway as a new target for mitigation of immunotherapy-associated intestinal inflammation. Given that they recapitulate both the phenotypic outcomes and underlying interlineage immune interactions, IIOs can be used to study tissue-resident immune responses in the context of tumorigenesis and infectious and autoimmune diseases.

DOI: https://doi.org/10.1038/s41586-024-07791-5
J Vis Exp. 2024 Jul 19.

Optimized Protocol for Intestinal Swiss Rolls and Immunofluorescent Staining of Paraffin Embedded Tissue.

Sarah A Dooley, Rachel Stubler, Rachel M Edens, Piper R McKee, Jordan N Rucker, Amy Engevik.

The intestine is a complex organ composed of the small and the large intestines. The small intestine can be further divided into duodenum, jejunum, and ileum. Each anatomical region of the intestine has a unique function that is reflected by differences in cellular structure. Investigating changes in the intestine requires an in-depth analysis of different tissue regions and cellular alterations. To study the intestine and visualize large pieces of tissue, researchers commonly use a technique known as intestinal Swiss rolls. In this technique, the intestine is divided into each anatomical region and fixed in a flat orientation. Then, the tissue is carefully rolled and processed for paraffin embedding. Proper tissue fixation and orientation is an often-overlooked laboratory technique but is critically important for downstream analysis. Additionally, improper Swiss rolling of intestinal tissue can damage the fragile intestinal epithelium, leading to poor tissue quality for immunostaining. Ensuring well-fixed and properly oriented tissue with intact cellular structures is a crucial step that ensures optimal visualization of intestinal cells. We present a cost-effective and simple method for making Swiss rolls to include all sections of the intestine in a single paraffin-embedded block. We also describe optimized immunofluorescence staining of intestinal tissue to study various aspects of the intestinal epithelium. The following protocol provides researchers with a comprehensive guide to obtaining high-quality immunofluorescence images through intestinal tissue fixation, Swiss-roll technique, and immunostaining. Employing these refined approaches preserves the intricate morphology of the intestinal epithelium and fosters a deeper understanding of intestinal physiology and pathobiology.

DOI: https://doi.org/10.3791/66977
Transl Cancer Res. 2024 Jul 31. 13(7): 3522-3535

The ratio of PKM1/PKM2 is the key factor affecting the glucose metabolism and biological function of colorectal cancer cells.

Liang Ma, Xue Zhang, Yan Liu, Hui Jin, Dan Li, Hui Zhang, Li Feng, Jing Zuo, Yudong Wang, Jiayin Liu, Jing Han.

   Background: Despite evidence suggesting a significant role of pyruvate kinase muscle isozyme (PKM) in cancer development, its particular function in colorectal cancer (CRC) remains unclear. This study aimed to elucidate the specific role and mechanism of PKM and its isoforms, PKM1 and PKM2, in the progression of CRC.
Methods: We analyzed PKM, PKM1, and PKM2 expression in CRC tissues and their correlation with clinicopathological features. Plasmids were constructed to modulate these isoforms' expression in CRC cells. Cellular behavior changes, including glucose metabolism alterations, were assessed using the Seahorse Energy Meter, and the Cell Counting Kit-8 (CCK8) assay to determine the inhibitory concentration of 5-fluorouracil (5-FU) on different CRC cell groups.
Results: Our results showed significant PKM overexpression in CRC tissues, which was correlated with negative prognostic factors such as advanced T stages and lymph node metastasis. A lower PKM1/PKM2 ratio was associated with these adverse outcomes. Functionally, PKM1 overexpression decreased cell migration and invasion, increasing 5-FU sensitivity. Conversely, PKM2 overexpression promoted malignant traits and reduced 5-FU sensitivity. Intriguingly, the introduction of glycolysis inhibitors attenuated the impact of PKM on the biological functions of CRC cells, suggesting a glycolysis-dependent mechanism.
Conclusions: This study establishes the PKM1/PKM2 ratio as crucial in CRC progression and 5-FU response. PKM1 overexpression reduces CRC malignancy and increases 5-FU sensitivity, while PKM2 does the opposite. Notably, glycolysis inhibitors lessen PKM's impact on CRC cells, highlighting a glycolysis-dependent mechanism. These insights suggest targeting PKM isoforms and glycolysis pathways as a promising CRC therapeutic strategy, potentially enhancing treatment efficacy.

Keywords:  Pyruvate kinase; chemotherapy sensitivity; clinicopathological features; glucose metabolism

DOI:  https://doi.org/10.21037/tcr-24-154
Cancer Cell. 2024 Aug 12. pii: S1535-6108(24)00271-X. [Epub ahead of print]42(8): 1370-1385.e9

CCL19-producing fibroblasts promote tertiary lymphoid structure formation enhancing anti-tumor IgG response in colorectal cancer liver metastasis.

Yifan Zhang, Guangjian Liu, Qianwen Zeng, Wenrui Wu, Kai Lei, Chuankai Zhang, Miaoling Tang, Yuting Zhang, Xiao Xiang, Li Tan, Rui Cui, Si Qin, Xinming Song, Changjun Yin, Zhihang Chen, Ming Kuang.

  Tertiary lymphoid structures (TLSs) are associated with enhanced immunity in tumors. However, their formation and functions in colorectal cancer liver metastasis (CRLM) remain unclear. Here, we reveal that intra- and peri-tumor mature TLSs (TLS+) are associated with improved clinical outcomes than TLS- tumors. Using single-cell-RNA-sequencing and spatial-enhanced-resolution-omics-sequencing (Stereo-seq), we reveal that TLS+ tumors are enriched with IgG+ plasma cells (PCs), while TLS- tumors are characterized with IgA+ PCs. By generating TLS-associated PC-derived monoclonal antibodies in vitro, we show that TLS-PCs secrete tumor-targeting antibodies. As the proof-of-concept, we demonstrate the anti-tumor activities of TLS-PC-mAb6 antibody in humanized mouse model of colorectal cancer. We identify a fibroblast lineage secreting CCL19 that facilitates lymphocyte trafficking to TLSs. CCL19 treatment promotes TLS neogenesis and prevents tumor growth in mice. Our data uncover the central role of CCL19+ fibroblasts in TLS formation, which in turn generates therapeutic antibodies to restrict CRLM.

Keywords:  CCL19; IgG plasma cell; Tertiary lymphoid structures; colorectal cancer liver metastasis; humanized mouse; monoclonal antibody; single-cell RNA sequencing; spatial transcriptomics

DOI:  https://doi.org/10.1016/j.ccell.2024.07.006
J Exp Clin Cancer Res. 2024 Aug 16. 43(1): 227

Palmitoyltransferase ZDHHC6 promotes colon tumorigenesis by targeting PPARγ-driven lipid biosynthesis via regulating lipidome metabolic reprogramming.

Junqi Shan, Xinyu Li, Runqi Sun, Yao Yao, Yan Sun, Qin Kuang, Xianling Dai, Yanlai Sun.

   BACKGROUND: The failure of proper recognition of the intricate nature of pathophysiology in colorectal cancer (CRC) has a substantial effect on the progress of developing novel medications and targeted therapy approaches. Imbalances in the processes of lipid oxidation and biosynthesis of fatty acids are significant risk factors for the development of CRC. Therapeutic intervention that specifically targets the peroxisome proliferator-activated receptor gamma (PPARγ) and its downstream response element, in response to lipid metabolism, has been found to promote the growth of tumors and has shown significant clinical advantages in cancer patients.
METHODS: Clinical CRC samples and extensive in vitro and in vivo experiments were carried out to determine the role of ZDHHC6 and its downstream targets via a series of biochemical assays, molecular analysis approaches and lipid metabolomics assay, etc. RESULTS: To study the effect of ZDHHC6 on the progression of CRC and identify whether ZDHHC6 is a palmitoyltransferase that regulates fatty acid synthesis, which directly palmitoylates and stabilizes PPARγ, and this stabilization in turn activates the ACLY transcription-related metabolic pathway. In this study, we demonstrate that PPARγ undergoes palmitoylation in its DNA binding domain (DBD) section. This lipid-related modification enhances the stability of PPARγ protein by preventing its destabilization. As a result, palmitoylated PPARγ inhibits its degradation induced by the lysosome and facilitates its translocation into the nucleus. In addition, we have identified zinc finger-aspartate-histidine-cysteine 6 (ZDHHC6) as a crucial controller of fatty acid biosynthesis. ZDHHC6 directly interacts with and adds palmitoyl groups to stabilize PPARγ at the Cys-313 site within the DBD domain of PPARγ. Consequently, this palmitoylation leads to an increase in the expression of ATP citrate lyase (ACLY). Furthermore, our findings reveals that ZDHHC6 actively stimulates the production of fatty acids and plays a role in the development of colorectal cancer. However, we have observed a significant reduction in the cancer-causing effects when the expression of ZDHHC6 is inhibited in in vivo trials. Significantly, in CRC, there is a strong positive correlation between the high expression of ZDHHC6 and the expression of PPARγ. Moreover, this high expression of ZDHHC6 is connected with the severity of CRC and is indicative of a poor prognosis.
CONCLUSIONS: We have discovered a mechanism in which lipid biosynthesis is controlled by ZDHHC6 and includes the signaling of PPARγ-ACLY in the advancement of CRC. This finding provides a justification for targeting lipid synthesis by blocking ZDHHC6 as a potential therapeutic approach.

Keywords:  Colorectal cancer (CRC); Lysosomal degradation; PPARγ; Palmitoylation; ZDHHC6

DOI:  https://doi.org/10.1186/s13046-024-03154-0
Trends Endocrinol Metab. 2024 Aug 08. pii: S1043-2760(24)00197-8. [Epub ahead of print]

Adapt and shape: metabolic features within the metastatic niche.

Erica Pranzini, Luigi Ippolito, Elisa Pardella, Elisa Giannoni, Paola Chiarugi.

  The success of disseminating cancer cells (DTCs) at specific metastatic sites is influenced by several metabolic factors. Even before DTCs arrival, metabolic conditioning from the primary tumor participates in creating a favorable premetastatic niche at distant organs. In addition, DTCs adjust their metabolism to better survive along the metastatic journey and successfully colonize their ultimate destination. However, the idea that the environment of the target organs may metabolically impact the metastatic fate is often underestimated. Here, we review the coexistence of two distinct strategies by which cancer cells shape and/or adapt to the metabolic profile of colonized tissues, ultimately creating a proper soil for their seeding and proliferation.

Keywords:  metabolic adaptation; metastatic niche; nutrient availability; organotropism; tissue metabolism

DOI:  https://doi.org/10.1016/j.tem.2024.07.016
FASEB J. 2024 Aug 31. 38(16): e70001

Interferon-gamma signaling drives epithelial TNF-alpha receptor-2 expression during colonic tissue repair.

Sean Watson, Rodolfo I Cabrera-Silva, Charles A Parkos, Asma Nusrat, Miguel Quiros.

  Interferon-gamma (IFNγ) is traditionally recognized for its pro-inflammatory role during intestinal inflammation. Here, we demonstrate that IFNγ also functions as a pro-repair molecule by increasing TNFα receptor 2 (TNFR2 protein/TNFRSF1B gene) expression on intestinal epithelial cells (IEC) following injury in vitro and in vivo. In silico analyses identified binding sites for the IFNγ signaling transcription factor STAT1 in the promoter region of TNFRSF1B. Scratch-wounded IEC exposed to IFNγ exhibited a STAT1-dependent increase in TNFR2 expression. In situ hybridization revealed elevated Tnfrsf1b mRNA levels in biopsy-induced colonic mucosal wounds, while intraperitoneal administration of IFNγ neutralizing antibodies following mucosal injury resulted in impaired IEC Tnfrsf1b mRNA and inhibited colonic mucosal repair. These findings challenge conventional notions that "pro-inflammatory" mediators solely exacerbate damage by highlighting latent pro-repair functions. Moreover, these results emphasize the critical importance of timing and amount in the synthesis and release of IFNγ and TNFα during the inflammatory process, as they are pivotal in restoring tissue homeostasis.

Keywords:  cytokines; inflammation; repair; wound healing

DOI:  https://doi.org/10.1096/fj.202401695
Nat Commun. 2024 Aug 13. 15(1): 6949

Dysfunctional tumor-infiltrating Vδ1 + T lymphocytes in microsatellite-stable colorectal cancer.

Victoria Stary, Ram V Pandey, Julia List, Lisa Kleissl, Florian Deckert, Julijan Kabiljo, Johannes Laengle, Vasileios Gerakopoulos, Rudolf Oehler, Lukas Watzke, Matthias Farlik, Samuel W Lukowski, Anne B Vogt, Georg Stary, Hannes Stockinger, Michael Bergmann, Nina Pilat.

Although γδ T cells are known to participate in immune dysregulation in solid tumors, their relevance to human microsatellite-stable (MSS) colorectal cancer (CRC) is still undefined. Here, using integrated gene expression analysis and T cell receptor sequencing, we characterized γδ T cells in MSS CRC, with a focus on Vδ1 + T cells. We identified Vδ1+ T cells with shared motifs in the third complementarity-determining region of the δ-chain, reflective of antigen recognition. Changes in gene and protein expression levels suggested a dysfunctional effector state of Vδ1+ T cells in MSS CRC, distinct from Vδ1+ T cells in microsatellite-instable (MSI). Interaction analysis highlighted an immunosuppressive role of fibroblasts in the dysregulation of Vδ1+ T cells in MSS CRC via the TIGIT-NECTIN2 axis. Blocking this pathway with a TIGIT antibody partially restored cytotoxicity of the dysfunctional Vδ1 phenotype. These results define an operative pathway in γδ T cells in MSS CRC.

DOI: https://doi.org/10.1038/s41467-024-51025-1
Trends Cancer. 2024 Aug 14. pii: S2405-8033(24)00146-8. [Epub ahead of print]

CFTR and colorectal cancer susceptibility: an urgent need for further studies.

S Spelier, S Derksen, R Hofland, J M Beekman, B Yetkin-Arik.

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene result in cystic fibrosis, a multiorgan disease characterized by aberrant epithelial cell fluid secretion. Recent studies describe a connection between CFTR malfunctioning and a heightened susceptibility to colorectal cancer (CRC). Here, we outline these links and suggest directions for further studies.

DOI: https://doi.org/10.1016/j.trecan.2024.07.006
Int J Mol Sci. 2024 Jul 30. pii: 8352. [Epub ahead of print]25(15):

The Role of Adipocytes Recruited as Part of Tumor Microenvironment in Promoting Colorectal Cancer Metastases.

Yunxia Ma, Miljana Nenkov, Yuan Chen, Nikolaus Gaßler.

  Adipose tissue dysfunction, which is associated with an increased risk of colorectal cancer (CRC), is a significant factor in the pathophysiology of obesity. Obesity-related inflammation and extracellular matrix (ECM) remodeling promote colorectal cancer metastasis (CRCM) by shaping the tumor microenvironment (TME). When CRC occurs, the metabolic symbiosis of tumor cells recruits adjacent adipocytes into the TME to supply energy. Meanwhile, abundant immune cells, from adipose tissue and blood, are recruited into the TME, which is stimulated by pro-inflammatory factors and triggers a chronic local pro-inflammatory TME. Dysregulated ECM proteins and cell surface adhesion molecules enhance ECM remodeling and further increase contractibility between tumor and stromal cells, which promotes epithelial-mesenchymal transition (EMT). EMT increases tumor migration and invasion into surrounding tissues or vessels and accelerates CRCM. Colorectal symbiotic microbiota also plays an important role in the promotion of CRCM. In this review, we provide adipose tissue and its contributions to CRC, with a special emphasis on the role of adipocytes, macrophages, neutrophils, T cells, ECM, and symbiotic gut microbiota in the progression of CRC and their contributions to the CRC microenvironment. We highlight the interactions between adipocytes and tumor cells, and potential therapeutic approaches to target these interactions.

Keywords:  CAAs; TME; adipocyte–mesenchymal transition; adipose tissue; colorectal cancer metastases; microbiota; obesity; therapeutics

DOI:  https://doi.org/10.3390/ijms25158352
Transl Cancer Res. 2024 Jul 31. 13(7): 3446-3464

SLC12A8 upregulation promotes colorectal cancer progression and chemoresistance.

Zhe Sun, Zhiyan Nie, Yao Xu, Yingshun Cui, Wenjian Ma, Tongcun Zhang.

   Background: Colorectal cancer (CRC), a prevalent gastrointestinal malignant disease, causes substantial morbidity and mortality. Identification of novel prognostic biomarkers and therapeutic targets is critically needed to improve patient outcomes. Although solute carrier family 12 member 8 (SLC12A8) has high expression in various tumors and affects tumor progression, its role in CRC remains unclear. The aim of this study was to investigate the functions of SLC12A8 in CRC.
Methods: SLC12A8 expression and its association with clinical significance in CRC patients were explored via multiple public databases, including The Cancer Genome Atlas (TCGA), Genotype-Tissue Expression (GTEx), The Human Protein Atlas (HPA), The University of ALabama at Birmingham CANcer data analysis Portal (UALCAN), and Kaplan-Meier plotter. The effects of SLC12A8 on the CRC cell apoptosis, epithelial-mesenchymal transition (EMT), reactive oxygen species (ROS) production, and sensitivity to oxaliplatin were verified by in vitro experiments.
Results: SLC12A8 expression was upregulated in CRC tissues compared with normal colorectal tissues. Furthermore, high expression of SLC12A8 was associated with poorer prognosis in CRC patients. Pathway enrichment analyses revealed SLC12A8 involvement in oxidative stress and transforming growth factor-beta (TGF-β) signaling. Experiments in CRC cells showed that SLC12A8 upregulation promoted apoptosis resistance, EMT, and inhibited ROS production. Moreover, SLC12A8 knockdown enhanced the sensitivity of CRC cells to oxaliplatin chemotherapy.
Conclusions: Our integrative analyses identify SLC12A8 as a candidate biomarker for CRC progression. Targeting SLC12A8 may improve patient responses to oxaliplatin-based treatment regimens.

Keywords:  Solute carrier family 12 member 8 (SLC12A8); colorectal cancer (CRC); oxaliplatin; reactive oxygen species (ROS)

DOI:  https://doi.org/10.21037/tcr-24-87
Cell Rep. 2024 Aug 14. pii: S2211-1247(24)00972-0. [Epub ahead of print]43(8): 114622

RPL22 is a tumor suppressor in MSI-high cancers and a splicing regulator of MDM4.

Hannah N W Weinstein, Kevin Hu, Lisa Fish, Yih-An Chen, Paul Allegakoen, Julia H Pham, Keliana S F Hui, Chih-Hao Chang, Meltem Tutar, Lorena Benitez-Rivera, Maria B Baco, Hanbing Song, Andrew O Giacomelli, Francisca Vazquez, Mahmoud Ghandi, Hani Goodarzi, Franklin W Huang.

  Microsatellite instability-high (MSI-H) tumors are malignant tumors that, despite harboring a high mutational burden, often have intact TP53. One of the most frequent mutations in MSI-H tumors is a frameshift mutation in RPL22, a ribosomal protein. Here, we identified RPL22 as a modulator of MDM4 splicing through an alternative splicing switch in exon 6. RPL22 loss increases MDM4 exon 6 inclusion and cell proliferation and augments resistance to the MDM inhibitor Nutlin-3a. RPL22 represses the expression of its paralog, RPL22L1, by mediating the splicing of a cryptic exon corresponding to a truncated transcript. Therefore, damaging mutations in RPL22 drive oncogenic MDM4 induction and reveal a common splicing circuit in MSI-H tumors that may inform therapeutic targeting of the MDM4-p53 axis and oncogenic RPL22L1 induction.

Keywords:  CP: Cancer; CP: Molecular biology; MDM4; MDM4 exon 6 inclusion; MSI-H; RPL22; RPL22 p.K15fs; alternative splicing; microsattelite instability-high; p53; ribosomal proteins; tumor suppressor

DOI:  https://doi.org/10.1016/j.celrep.2024.114622
Cancer Cell. 2024 Aug 12. pii: S1535-6108(24)00270-8. [Epub ahead of print]42(8): 1386-1400.e8

Integrative plasma and fecal metabolomics identify functional metabolites in adenoma-colorectal cancer progression and as early diagnostic biomarkers.

Yang Sun, Xiang Zhang, Dong Hang, Harry Cheuk-Hay Lau, Jie Du, Chuanfa Liu, Mingxu Xie, Yasi Pan, Le Wang, Cong Liang, Xingyu Zhou, Danyu Chen, Jiamei Rong, Zengren Zhao, Alvin Ho-Kwan Cheung, Yuet Wu, Hongyan Gou, Chi Chun Wong, Lingbin Du, Junliang Deng, Zhibin Hu, Hongbing Shen, Yinglei Miao, Jun Yu.

  Changes in plasma and fecal metabolomes in colorectal cancer (CRC) progression (normal-adenoma-CRC) remain unclear. Here, plasma and fecal samples were collected from four independent cohorts of 1,251 individuals (422 CRC, 399 colorectal adenoma [CRA], and 430 normal controls [NC]). By metabolomic profiling, signature plasma and fecal metabolites with consistent shift across NC, CRA, and CRC are identified, including CRC-enriched oleic acid and CRC-depleted allocholic acid. Oleic acid exhibits pro-tumorigenic effects in CRC cells, patient-derived organoids, and two murine CRC models, whereas allocholic acid has opposing effects. By integrative analysis, we found that oleic acid or allocholic acid directly binds to α-enolase or farnesoid X receptor-1 in CRC cells, respectively, to modulate cancer-associated pathways. Clinically, we establish a panel of 17 plasma metabolites that accurately diagnoses CRC in a discovery and three validation cohorts (AUC = 0.848-0.987). Overall, we characterize metabolite signatures, mechanistic significance, and diagnostic potential of plasma and fecal metabolomes in CRC.

Keywords:  biomarkers; colorectal adenoma; colorectal cancer; cytokines; fecal metabolites; integrative analysis; microbiota; non-invasive diagnosis; plasma metabolites

DOI:  https://doi.org/10.1016/j.ccell.2024.07.005
Front Cell Dev Biol. 2024 ;12 1375543

The role of cancer-associated fibroblasts in the invasion and metastasis of colorectal cancer.

Jinjin Yin, Wenting Zhu, Senling Feng, Pengke Yan, Shumin Qin.

  Colorectal cancer (CRC) is the third most common cancer and has ranked the third leading cause in cancerassociated death globally. Metastasis is the leading cause of death in colorectal cancer patients. The role of tumor microenvironment (TME) in colorectal cancer metastasis has received increasing attention. As the most abundant cell type in the TME of solid tumors, cancer-associated fibroblasts (CAFs) have been demonstrated to have multiple functions in advancing tumor growth and metastasis. They can remodel the extracellular matrix (ECM) architecture, promote epithelial-mesenchymal transition (EMT), and interact with cancer cells or other stromal cells by secreting growth factors, cytokines, chemokines, and exosomes, facilitating tumor cell invasion into TME and contributing to distant metastasis. This article aims to analyze the sources and heterogeneity of CAFs in CRC, as well as their role in invasion and metastasis, in order to provide new insights into the metastasis mechanism of CRC and its clinical applications.

Keywords:  cancer-associated fibroblasts; colorectal cancer; heterogeneity; invasion; metastasis

DOI:  https://doi.org/10.3389/fcell.2024.1375543