Whole-mount staining of mouse colorectal cancer organoids and fibroblast-organoid co-cultures

Summary: Imaging organoid culture provides an excellent tool for studying complex diseases such as cancer. However, retaining the morphology of intact organoids for immunolabeling has been challenging. Here, we describe a protocol for immunofluorescence staining in intact colorectal cancer organoids derived from mice. We also describe additional steps for co-culture with mouse fibroblasts to enable the study of interactions with other cellular components of the tissue microenvironment.For complete details on the use and execution of this protocol, please refer to Martinez-Ordoñez et al. (2023).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics

THBS1-producing tumor-infiltrating monocyte-like cells contribute to immunosuppression and metastasis in colorectal cancer

Abstract Mesenchymal activation, characterized by dense stromal infiltration of immune and mesenchymal cells, fuels the aggressiveness of colorectal cancers (CRC), driving progression and metastasis. Targetable molecules in the tumor microenvironment (TME) need to be identified to improve the outcome in CRC patients with this aggressive phenotype. This study reports a positive link between high thrombospondin-1 (THBS1) expression and mesenchymal characteristics, immunosuppression, and unfavorable CRC prognosis. Bone marrow-derived monocyte-like cells recruited by CXCL12 are the primary source of THBS1, which contributes to the development of metastasis by inducing cytotoxic T-cell exhaustion and impairing vascularization. Furthermore, in orthotopically generated CRC models in male mice, THBS1 loss in the TME renders tumors partially sensitive to immune checkpoint inhibitors and anti-cancer drugs. Our study establishes THBS1 as a potential biomarker for identifying mesenchymal CRC and as a critical suppressor of antitumor immunity that contributes to the progression of this malignancy with a poor prognosis

Hydrogenation of CuBTC Framework with the Introduction of a PtC Hydrogen Spillover Catalyst

Hydrogen uptake of a microporous metal organic framework, CuBTC, is increased 3.5-fold at 298 K and 20 bar upon the addition of a hydrogen spillover catalyst, from 0.17 to 0.61 wt %. Structural integrity upon mixing with the catalyst is important to achieve this level of uptake. Increasing the adsorption temperature to 323 K significantly reduces the rate of uptake, but 0.55 wt % uptake is observed when the experimental equilibration time is extended. The slow, pressure-independent uptake at 323 K, along with the desorption behavior is suggestive of a hydrogenation process of the CuBTC substrate. PXRD analysis suggests the hydrogenated sample remains intact and FTIR demonstrates hydrogenation of the carboxylate group of the BTC ligand but finds no evidence for hydrogenation of the carbons of the BTC ligand. Although hydrogenation of the CuBTC does not lead to readily desorbable H2, the results shed light on a possible mechanism of the hydrogen spillover process

THBS1-producing tumor-infiltrating monocyte-like cells contribute to immunosuppression and metastasis in colorectal cancer

Mesenchymal activation, characterized by dense stromal infiltration of immune and mesenchymal cells, fuels the aggressiveness of colorectal cancers (CRC), driving progression and metastasis. Targetable molecules in the tumor microenvironment (TME) need to be identified to improve the outcome in CRC patients with this aggressive phenotype. This study reports a positive link between high thrombospondin-1 (THBS1) expression and mesenchymal characteristics, immunosuppression, and unfavorable CRC prognosis. Bone marrow-derived monocyte-like cells recruited by CXCL12 are the primary source of THBS1, which contributes to the development of metastasis by inducing cytotoxic T-cell exhaustion and impairing vascularization. Furthermore, in orthotopically generated CRC models in male mice, THBS1 loss in the TME renders tumors partially sensitive to immune checkpoint inhibitors and anti-cancer drugs. Our study establishes THBS1 as a potential biomarker for identifying mesenchymal CRC and as a critical suppressor of antitumor immunity that contributes to the progression of this malignancy with a poor prognosis.大腸がん：骨髄が転移を促進？ --新しい治療法への展望--. 京都大学プレスリリース. 2023-10-27