bims-instec 2023-07-30 papers

bims-instec

Biomed News

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

Issue of 2023–07–30
eight papers selected by
Maria-Virginia Giolito, Université Catholique de Louvain

Sonic Hedgehog and WNT signaling regulate a positive feedback loop between intestinal epithelial and stromal cells to promote epithelial regeneration.
Colorectal cancer organoid-stroma biobank allows subtype-specific assessment of individualized therapy responses.
Integrin α7β1 represses intestinal absorptive cell differentiation.
Mice lacking intestinal Nr1i2 have normal intestinal homeostasis under steady-state conditions and are not hypersensitive to inflammation under lipopolysaccharide treatment.
Transcriptional Profiling and Consensus Molecular Subtype Assignment to Understand Response and Resistance to Anti-Epidermal Growth Factor Receptor Therapy in Colorectal Cancer.
Mammalian Intestinal Development and Differentiation - the State of the Art.
Formyl peptide receptor-1 (FPR1) represses intestinal oncogenesis.
Comparative analyses of the prognosis, tumor immune microenvironment, and drug treatment response between left-sided and right-sided colon cancer by integrating scRNA-seq and bulk RNA-seq data.

Cell Mol Gastroenterol Hepatol. 2023 Jul 20. pii: S2352-345X(23)00131-5. [Epub ahead of print]

Sonic Hedgehog and WNT signaling regulate a positive feedback loop between intestinal epithelial and stromal cells to promote epithelial regeneration.

Emilia J Orzechowska-Licari, Agnieszka B Bialkowska, Vincent W Yang.

   BACKGROUND AND AIMS: Active intestinal stem cells in the intestinal epithelium are prone to injury by ionizing radiation. We previously showed that upon radiation-induced injury, normally quiescent reserve ISCs (rISCs) marked by B-cell-specific Moloney murine leukemia virus integration site 1 (BMI1) are activated by MUSASHI1 (MSI1) and exit from the quiescent state to regenerate the intestinal epithelium. This study aims to further establish the mechanism that regulates activation of Bmi1-CreER;Rosa26eYFP (Bmi1-CreER) rISCs following γ radiation-induced injury.
METHODS: Bmi1-CreER mice were treated with tamoxifen to initiate lineage tracing of BMI1 (eYFP+) cells and exposed to 12 Gy of total body γ irradiation (TBI) or sham. Intestinal tissues were collected and analyzed by immunofluorescence, Western blot, qRT-PCR, ELISA, and ChIP-PCR.
RESULTS: After irradiation, increased expression of Msi1 in eYFP+ cells was accompanied by increased expression of Axin2, a WNT marker. Promoter studies of the Msi1 gene indicated that Msi1 is a WNT target gene. Co-culture of stromal cells isolated from irradiated mice stimulated Bmi1-CreER-derived organoids regeneration more effectively than those from sham mice. Expression of WNT ligands, including Wnt2b, Wnt4, Wnt5a, and Rspo3, was increased in irradiated stromal cells compared to sham-treated stromal cells. Moreover, expression of the Sonic Hedgehog (SHH) effector, Gli1, was increased in stromal cells from irradiated mice. This was correlated with an increased expression of SHH in epithelial cells post-irradiation, indicating epithelial-stromal interaction. Finally, pre-injury treatment with SHH inhibitor cyclopamine significantly reduced intestinal epithelial regeneration and Msi1 expression post-irradiation.
CONCLUSIONS: Upon ionizing radiation-induced injury, intestinal epithelial cells increase SHH secretion, stimulating stromal cells to secrete WNT ligands. WNT activators induce Msi1 expression in the Bmi1-CreER cells. This stromal-epithelial interaction leads to Bmi1-CreER rISCs induction and epithelial regeneration.

Keywords:  GLI1; MSI1; Sonic Hedgehog; WNT signaling; intestinal epithelium; regeneration

DOI:  https://doi.org/10.1016/j.jcmgh.2023.07.004
Cancer Discov. 2023 Jul 25. pii: CD-23-0050. [Epub ahead of print]

Colorectal cancer organoid-stroma biobank allows subtype-specific assessment of individualized therapy responses.

Henner F Farin, Mohammed H Mosa, Benardina Ndreshkjana, Britta M Grebbin, Birgit Ritter, Constantin Menche, Kilian B Kennel, Paul K Ziegler, Lili Szabo, Julia Bollrath, Dietmar Rieder, Birgitta E Michels, Alena Kress, Muge Bozlar, Tahmineh Darvishi, Sara Stier, Ivan-Maximiliano Kur, Katrin Bankov, Rebecca Kesselring, Stefan Fichtner-Feigl, Bernhard Brune, Thorsten O Goetze, Salah-Eddin Al-Batran, Christian H Brandts, Wolf O Bechstein, Peter J Wild, Andreas Weigert, Susanne Muller, Stefan Knapp, Zlatko Trajanoski, Florian R Greten.

In colorectal cancers (CRC) the tumor microenvironment plays a key role for prognosis and therapy efficacy. Patient-derived tumor organoids (PDTOs) show enormous potential for preclinical testing, however, cultured tumor cells lose important characteristics including the 'consensus molecular subtypes' (CMS). To better reflect the cellular heterogeneity, we established the CRC organoid-stroma biobank of matched PDTOs and cancer-associated fibroblasts (CAFs) from 30 patients. Context-specific phenotyping showed that xenotransplantation or co-culture with CAFs improves the transcriptomic fidelity and instructs subtype-specific stromal gene expression. Furthermore, functional profiling in co-culture exposed CMS4-specific therapeutic resistance to Gefitinib and SN-38 and prognostic expression signatures. Chemogenomic library screening identified patient- and therapy-dependent mechanisms of stromal resistance including MET as common target. Our results demonstrate that CRC phenotypes are encrypted in the cancer epithelium in a plastic fashion that strongly depends on the context. Consequently, CAFs are essential for faithful representation of molecular subtypes and therapy responses ex vivo.

DOI: https://doi.org/10.1158/2159-8290.CD-23-0050
Exp Cell Res. 2023 Jul 24. pii: S0014-4827(23)00271-9. [Epub ahead of print] 113723

Integrin α7β1 represses intestinal absorptive cell differentiation.

Gabriel Cloutier, Amira Seltana, Sepideh Fallah, Jean-François Beaulieu.

  Intestinal epithelial cell differentiation is a highly controlled and orderly process occurring in the crypt so that cells migrating out to cover the villi are already fully functional. Absorptive cell precursors, which originate from the stem cell population located in the lower third of the crypt, are subject to several cycles of amplification in the transit amplifying (TA) zone, before reaching the terminal differentiation compartment located in the upper third. There is a large body of evidence that absorptive cell differentiation is halted in the TA zone through various epigenetic, transcriptional and intracellular signalling events or mechanisms allowing the transient expansion of this cell population but how these mechanisms are themself regulated remains obscure. One clue can be found in the epithelial cell-matrix microenvironment located all along the crypt-villus axis. Indeed, a previous study from our group revealed that α5-subunit containing laminins such as lamimin-511 and 512 inhibit early stages of differentiation in Caco-2/15 cells. Among potential receptors for laminin 511/512 is the integrin α7β1, which has previously been reported to be expressed in the human intestinal crypts and in early stages of Caco-2/15 cell differentiation. In this study, the effects of knocking down ITGA7 in Caco-2/15 cells were studied using shRNA and CRISPR/Cas9 strategies. Abolition of the α7 integrin subunit resulted in a significant increase in the level of differentiation and polarization markers as well as the morphological features of intestinal cells. Activities of focal adhesion kinase and Src kinase were both reduced in α7-knockdown cells while the three major intestinal pro-differentiation factors CDX2, HNFα1 and HNF4α were overexpressed. Two epigenetic events associated with intestinal differentiation, the reduction of tri-methylated lysine 27 on histone H3 and the increase of acetylation of histone H4 were also observed in α7-knockdown cells. On the other hand, the ablation of α7 had no effect on cell proliferation. In conclusion, these data indicate that integrin α7β1 acts as a major repressor of absorptive cell terminal differentiation in the Caco-2/15 cell model and suggest that the laminin-α7β1 integrin interaction occurring in the transit amplifying zone of the adult intestine is involved in the transient halting of absorptive cell terminal differentiation.

Keywords:  Absorptive cell; Cell-matrix interactions; Differentiation; Integrin; Intestine

DOI:  https://doi.org/10.1016/j.yexcr.2023.113723
FASEB J. 2023 08;37(8): e23117

Mice lacking intestinal Nr1i2 have normal intestinal homeostasis under steady-state conditions and are not hypersensitive to inflammation under lipopolysaccharide treatment.

Salah Edden Amini, Sophie Emilie Bresson, Jérôme Ruzzin.

  Nr1i2, a nuclear receptor known for its key function in xenobiotic detoxification, has emerged as a potential regulator of intestinal homeostasis and inflammation. However, the role of Nr1i2 in different intestinal segments remains poorly known. Moreover, in vivo investigations on intestinal Nr1i2 have essentially been performed in whole-body Nr1i2 knockout (Nr1i2-/- ) mice where the deletion of Nr1i2 in all tissues may affect the intestinal phenotype. To better understand the role of Nr1i2 in the intestine, we generated intestinal epithelial-specific Nr1i2 knockout (iNr1i2-/- ) mice and studied the duodenum, jejunum, ileum, and colon of these animals during steady-state conditions and lipopolysaccharide (LPS)-induced inflammation. As compared to control (iNr1i2+/+ ) mice, iNr1i2-/- mice showed normal intestinal permeability as assessed by in vivo FITC-dextran test. The expression of genes involved in inflammation, tight- and adherens-junction, proliferation, glucose, and lipid metabolism was comparable in the duodenum, jejunum, ileum, and colon of iNr1i2-/- and iNr1i2+/+ mice. In line with these findings, histological analyses of the jejunum revealed no difference between iNr1i2-/- and iNr1i2+/+ mice. When treated with LPS, the intestine of iNr1i2-/- mice had no increased inflammatory response as compared to iNr1i2+/+ mice. Moreover, the health monitoring of LPS-treated iNr1i2-/- and iNr1i2+/+ mice was similar. Taken together, our results demonstrate that the specific deletion of Nr1i2 in the intestinal epithelium does not cause major intestinal damages in mice during both steady-state and inflammatory conditions.

Keywords:  LPS; environmental nuclear receptor; inflammation; intestine; transcription factors; transgenic mice

DOI:  https://doi.org/10.1096/fj.202301126
JCO Precis Oncol. 2023 Jul;7 e2200422

Transcriptional Profiling and Consensus Molecular Subtype Assignment to Understand Response and Resistance to Anti-Epidermal Growth Factor Receptor Therapy in Colorectal Cancer.

Saikat Chowdhury, Ria Gupta, Joshua Millstein, Kangyu Lin, Valsala Haridas, Mohammad A Zeineddine, Christine Parseghian, Heinz-Josef Lenz, Scott Kopetz, John Paul Shen.

PURPOSE: Activating mutations in KRAS, NRAS, and BRAF are known to cause resistance to anti-epidermal growth factor receptor (EGFR) therapy; however, only approximately 40% of patients with colorectal cancer (CRC) with RASWT tumors respond to anti-EGFR treatment. We sought to discover novel biomarkers to predict response to anti-EGFR antibody treatment in CRC and to understand mechanisms of resistance to anti-EGFR therapy.
MATERIALS AND METHODS: Transcriptomic profiles from three clinical and two preclinical cohorts treated with cetuximab were used to assign consensus molecular subtypes (CMS) to each sample and correlated with outcomes.
RESULTS: Restricting to RASWT patients, we observed that CMS2 tumors (canonical subtype) had significantly higher response rates relative to other CMS when treated with cetuximab combination with doublet chemotherapy (Okita et al cohort: 92% disease control rate (DCR) for CMS2, chi-square P = .04; CALGB/SWOG 80405 cohort: 90% objective response rate (ORR) for CMS2, chi-square P < .001) and with single-agent cetuximab (68%, chi-square P = .01). CMS2 tumors showed best response among right-sided (ORR = 80%) and left-sided (ORR = 92%) tumors in the CALGB/SWOG 80405 cohort. CMS2 cells lines were most likely to be sensitive to cetuximab (60%) and CMS2 patient-derived xenograft had the highest DCR (84%). We found Myc, E2F, and mammalian target of rapamycin pathways were consistently upregulated in resistant samples (enrichment score >1, false discovery rate <0.25). Inhibitors of these pathways in resistant cell lines exhibited additive effects with cetuximab.
CONCLUSION: These data suggest that CRC transcriptional profiles, when used to assign CMS, provide additional ability to predict response to anti-EGFR therapy relative to using tumor sidedness alone. Notably both right-sided and left-sided CMS2 tumors had excellent response, suggesting that anti-EGFR therapy be included as a treatment option for right-sided CMS2 tumors.

DOI: https://doi.org/10.1200/PO.22.00422
Cell Mol Gastroenterol Hepatol. 2023 Jul 26. pii: S2352-345X(23)00140-6. [Epub ahead of print]

Mammalian Intestinal Development and Differentiation - the State of the Art.

Hannah M Kolev, Klaus H Kaestner.

The development of the mammalian intestine from its earliest origins as a morphologically uniform sheet of endoderm cells during gastrulation into the complex organ system that is essential for the life of the organism is a truly fascinating process. During mid-gestation development, reciprocal interactions between endoderm-derived epithelium and mesoderm-derived mesenchyme enable villification, or the conversion of a radially symmetric pseudostratified epithelium into the functional subdivision of crypts and villi. Once a mature crypt-villus axis is established, proliferation and differentiation of new epithelial cells continue throughout life. Spatially localized signals including the Wnt, FGF, and Hippo systems among others ensure that new cells are continuously being born in the crypt. As cells exit the crypt compartment, a gradient of BMP signaling limits proliferation to allow for the specification of multiple mature cell types. The first major differentiation decision is dependent on Notch signaling, which specifies epithelial cells into absorptive and secretory lineages. The secretory lineage is further subdivided into Paneth, goblet, tuft and enteroendocrine cells via a complex network of transcription factors. While some of the signaling molecules are produced by epithelial cells, critical components are derived from specialized crypt-adjacent mesenchymal cells termed telocytes, which are marked by Foxl1, Gli1 and PDGFRα. The crucial nature of these processes is evidenced by the multitude of intestinal disorders such as colorectal cancer, short bowel syndrome, and inflammatory bowel disease which all reflect perturbations of the development and/or differentiation of the intestine.

DOI: https://doi.org/10.1016/j.jcmgh.2023.07.011
Oncoimmunology. 2023 ;12(1): 2237354

Formyl peptide receptor-1 (FPR1) represses intestinal oncogenesis.

Julie Le Naour, Léa Montégut, Yuhong Pan, Sarah Adriana Scuderi, Pierre Cordier, Adrien Joseph, Allan Sauvat, Valerio Iebba, Juliette Paillet, Gladys Ferrere, Ludivine Brechard, Claire Mulot, Grégory Dubourg, Laurence Zitvogel, Jonathan G Pol, Erika Vacchelli, Pierre-Laurent Puig, Guido Kroemer.

  Formyl peptide receptor-1 (FPR1) is a pattern recognition receptor that is mostly expressed by myeloid cells. In patients with colorectal cancer (CRC), a loss-of-function polymorphism (rs867228) in the gene coding for FPR1 has been associated with reduced responses to chemotherapy or chemoradiotherapy. Moreover, rs867228 is associated with accelerated esophageal and colorectal carcinogenesis. Here, we show that dendritic cells from Fpr1-/- mice exhibit reduced migration in response to chemotherapy-treated CRC cells. Moreover, Fpr1-/- mice are particularly susceptible to chronic ulcerative colitis and colorectal oncogenesis induced by the mutagen azoxymethane followed by oral dextran sodium sulfate, a detergent that induces colitis. These experiments were performed after initial co-housing of Fpr1-/- mice and wild-type controls, precluding major Fpr1-driven differences in the microbiota. Pharmacological inhibition of Fpr1 by cyclosporin H also tended to increase intestinal oncogenesis in mice bearing the ApcMin mutation, and this effect was reversed by the anti-inflammatory drug sulindac. We conclude that defective FPR1 signaling favors intestinal tumorigenesis through the modulation of the innate inflammatory/immune response.

Keywords:  Bone marrow-derived dendritic cells; chemotaxis; immunogenic chemotherapy; immunosurveillance; tumor microenvironment

DOI:  https://doi.org/10.1080/2162402X.2023.2237354
Aging (Albany NY). 2023 Jul 21. 15

Comparative analyses of the prognosis, tumor immune microenvironment, and drug treatment response between left-sided and right-sided colon cancer by integrating scRNA-seq and bulk RNA-seq data.

Lichao Cao, Shenrui Zhang, Danni Yao, Ying Ba, Qi Weng, Jin Yang, Hezi Zhang, Yanan Ren.

   BACKGROUND: In this study, we compared the prognosis, tumor immune microenvironment (TIM), and drug treatment response between left-sided (LCC) and right-sided (RCC) colon cancer to predict outcomes in patients with LCC and RCC.
METHODS: Based on identified differentially expressed genes and using single-cell RNA sequencing data, we constructed and validated a prognostic model for LCC and RCC patients in the TCGA-COAD cohort and GSE103479 cohort. Moreover, we compared the differences of TIM characteristics and drug treatment response between LCC and RCC patients.
RESULTS: We constructed and validated a five-gene prognostic model for LCC patients and a four-gene prognostic model for RCC patients, and both showed excellent performance. The RCC patients with higher risk scores were significantly associated with greater metastasis (P = 2.6×10-5), N stage (P = 0.012), advanced pathological stage (P = 1.4×10-4), and more stable microsatellite status (P = 0.007) but not T stage (P = 0.200). For LCC patients, the risk scores were not significantly associated with tumor stage and microsatellite status (P > 0.05). Additionally, immune infiltration by CD8 and regulatory T cells and M0, M1, and M2 macrophages differed significantly between LCC and RCC patients (P < 0.05). APC and TP53 mutations were significantly more common in LCC patients (P < 0.05). In contrast, KRAS, SYNE1, and MUC16 mutations were significantly more common in RCC patients (P < 0.05). In addition, tumor mutation burden values were significantly higher in RCC patients than in LCC patients (P = 5.9×10-8). Moreover, the expression of immune checkpoint targets was significantly higher in RCC patients than in LCC patients (P < 0.05), indicating that RCC patients maybe more sensitive to immunotherapy. However, LCC and RCC patients did not differ significantly in their sensitivity to eight selected chemicals or target drugs (P > 0.05). The average half-maximal inhibitory concentrations for camptothecin, teniposide, vinorelbine, and mitoxantrone were significantly lower in low-risk than in high-risk RCC patients (P < 0.05), indicating that the lower risk score of RCC patients, the more sensitive they were to these four drugs.
CONCLUSIONS: We investigated the differences in prognosis, TIM, and drug treatment response between LCC and RCC patients, which may contribute to accurate colon cancer prognosis and treatment of colon cancer.

Keywords:  colon cancer; left-sided and right-sided; prognosis; scRNA-seq; treatment

DOI:  https://doi.org/10.18632/aging.204894