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Gut Aug 2023The role of N-methyladenosine (mA) in tumour immune microenvironment (TIME) remains understudied. Here, we elucidate function and mechanism of YTH N-methyladenosine RNA...
OBJECTIVE
The role of N-methyladenosine (mA) in tumour immune microenvironment (TIME) remains understudied. Here, we elucidate function and mechanism of YTH N-methyladenosine RNA binding protein 1 (YTHDF1) in colorectal cancer (CRC) TIME.
DESIGN
Clinical significance of YTHDF1 was assessed in tissue microarrays (N=408) and TCGA (N=526) cohorts. function was determined in syngeneic tumours, intestine-specific knockin mice, and humanised mice. Single-cell RNA-seq (scRNA-seq) was employed to profile TIME. Methylated RNA immunoprecipitation sequencing (MeRIP-seq), RNA sequencing (RNA-seq) and ribosome sequencing (Ribo-seq) were used to identify YTHDF1 direct targets. Vesicle-like nanoparticles (VNPs)-encapsulated -siRNA was used for silencing in vivo.
RESULTS
expression negatively correlated with interferon-γ gene signature in TCGA-CRC. Concordantly, YTHDF1 protein negatively correlated with CD8 T-cell infiltration in independent tissue microarrays cohorts, implying its role in TIME. Genetic depletion of augmented antitumour immunity in CT26 (MSS-CRC) and MC38 (MSI-H-CRC) syngeneic tumours, while knockin promoted an immunosuppressive TIME facilitating CRC in azoxymethane-dextran sulphate-sodium or models. scRNA-seq identified reduction of myeloid-derived suppressor cells (MDSCs), concomitant with increased cytotoxic T cells in knockout tumours. Integrated MeRIP-seq, RNA-seq and Ribo-seq revealed p65/Rela as a YTHDF1 target. YTHDF1 promoted p65 translation to upregulate CXCL1, which increased MDSC migration via CXCL1-CXCR2 axis. Increased MSDCs in turn antagonised functional CD8 T cells in TIME. Importantly, targeting YTHDF1 by CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) or VNPs-si boosted anti-PD1 efficacy in MSI-H CRC, and overcame anti-PD1 resistance in MSS CRC.
CONCLUSION
YTHDF1 impairs antitumour immunity via an mA-p65-CXCL1/CXCR2 axis to promote CRC and serves as a therapeutic target in immune checkpoint blockade therapy.
Topics: Mice; Animals; CD8-Positive T-Lymphocytes; Colonic Neoplasms; Colorectal Neoplasms; Tumor Microenvironment
PubMed: 36717220
DOI: 10.1136/gutjnl-2022-328845 -
Gut Nov 2023is a probiotic species that can suppress intestinal inflammation by producing metabolites. We aimed to study the role of in colorectal tumourigenesis and immunotherapy.
OBJECTIVE
is a probiotic species that can suppress intestinal inflammation by producing metabolites. We aimed to study the role of in colorectal tumourigenesis and immunotherapy.
DESIGN
abundance was evaluated in stools of patients with colorectal cancer (CRC) (n=444) and healthy controls (n=575). The effects of were studied in or azoxymethane (AOM)-induced CRC mouse models, and in syngeneic mouse xenograft models of CT26 (microsatellite instability (MSI)-low) or MC38 (MSI-high). The change of immune landscape was evaluated by multicolour flow cytometry and immunohistochemistry staining. Metabolites were profiled by metabolomic profiling.
RESULTS
was significantly depleted in stools of patients with CRC compared with healthy controls. administration significantly inhibited tumour formation in mice, which was confirmed in mice with AOM-induced CRC. restored gut barrier function as indicated by improved intestinal permeability and enhanced expression of tight junction proteins. Butyrate was identified as the functional metabolite generated by or butyrate suppressed tumour growth by inducing cytotoxic granzyme B, interferon (IFN)-γ and tumour necrosis factor (TNF)-α CD8 T cells in orthotopic mouse models of MC38 or CT26. or butyrate also significantly improved antiprogrammed cell death protein 1 (anti-PD-1) efficacy in mice bearing MSI-low CT26 tumours. Mechanistically, butyrate directly bound to toll-like receptor 5 (TLR5) receptor on CD8 T cells to induce its activity through activating nuclear factor kappa B (NF-κB) signalling.
CONCLUSION
protects against colorectal tumourigenesis by producing butyrate, which could also improve anti-PD-1 efficacy by inducing functional CD8 T cells. is a potential adjuvant to augment anti-PD-1 efficacy against CRC.
Topics: Humans; Mice; Animals; CD8-Positive T-Lymphocytes; Butyrates; Carcinogenesis; Cell Transformation, Neoplastic; Colorectal Neoplasms
PubMed: 37491158
DOI: 10.1136/gutjnl-2023-330291 -
Cell Feb 2024Human brain development involves an orchestrated, massive neural progenitor expansion while a multi-cellular tissue architecture is established. Continuously expanding...
Human brain development involves an orchestrated, massive neural progenitor expansion while a multi-cellular tissue architecture is established. Continuously expanding organoids can be grown directly from multiple somatic tissues, yet to date, brain organoids can solely be established from pluripotent stem cells. Here, we show that healthy human fetal brain in vitro self-organizes into organoids (FeBOs), phenocopying aspects of in vivo cellular heterogeneity and complex organization. FeBOs can be expanded over long time periods. FeBO growth requires maintenance of tissue integrity, which ensures production of a tissue-like extracellular matrix (ECM) niche, ultimately endowing FeBO expansion. FeBO lines derived from different areas of the central nervous system (CNS), including dorsal and ventral forebrain, preserve their regional identity and allow to probe aspects of positional identity. Using CRISPR-Cas9, we showcase the generation of syngeneic mutant FeBO lines for the study of brain cancer. Taken together, FeBOs constitute a complementary CNS organoid platform.
Topics: Humans; Brain; Central Nervous System; Extracellular Matrix; Organoids; Pluripotent Stem Cells; Prosencephalon; Tissue Culture Techniques; Stem Cells; Morphogenesis
PubMed: 38194967
DOI: 10.1016/j.cell.2023.12.012 -
Advanced Science (Weinheim,... Jul 2023Unveiling biomedical functions of tumor-resident microbiota is challenging for developing advanced anticancer medicines. This study demonstrates that isolated...
Unveiling biomedical functions of tumor-resident microbiota is challenging for developing advanced anticancer medicines. This study demonstrates that isolated intratumoral bacteria, associated with natural purple photosynthetic bacteria, have inherent biocompatibility and strong immunogenic anticancer efficacies. They preferentially grow and proliferate within a targeted tumor milieu, which effectively causes immune cells to infiltrate the tumor and provoke strong anticancer responses in various syngeneic mouse models, including colorectal cancer, sarcoma, metastatic lung cancer, and extensive drug-resistant breast cancer. Furthermore, these functional bacteria-treated mice exhibit excellent anticancerous responses and have significantly prolonged survival rates with effective immunological memory. Light-harvesting nanocomplexes of microbial consortia of intratumoral bacteria and purple photosynthetic bacteria can diagnose tumors using bio-optical-window near-infrared light, making them useful theranostic agents for highly targeted immunological elimination of the tumor and for precisely marking tumor location.
Topics: Mice; Animals; Precision Medicine; Antineoplastic Agents; Disease Models, Animal; Lung Neoplasms; Bacteria
PubMed: 37150857
DOI: 10.1002/advs.202301679 -
Nature Communications Jul 2023The often immune-suppressive tumor microenvironment (TME) may hinder immune evasion and response to checkpoint blockade therapies. Pharmacological activation of the...
The often immune-suppressive tumor microenvironment (TME) may hinder immune evasion and response to checkpoint blockade therapies. Pharmacological activation of the STING pathway does create an immunologically hot TME, however, systemic delivery might lead to undesired off-target inflammatory responses. Here, we generate a small panel of esterase-activatable pro-drugs based on the structure of the non-nucleotide STING agonist MSA-2 that are subsequently stably incorporated into a liposomal vesicle for intravenous administration. The pharmacokinetic properties and immune stimulatory capacity of pro-drugs delivered via liposomes (SAProsomes) are enhanced compared to the free drug form. By performing efficacy screening among the SAProsomes incorporating different pro-drugs in syngeneic mouse tumor models, we find that superior therapeutic performance relies on improved delivery to the desired tumor and lymphoid compartments. The best candidate, SAProsome-3, highly stimulates secretion of inflammatory cytokines and creates a tumoricidal immune landscape. Notably, upon application to breast cancer or melanoma mouse models, SAProsome-3 elicits durable remission of established tumors and postsurgical tumor-free survival while decreasing metastatic burden without significant systemic toxicity. In summary, our work establishes the proof of principle for a better targeted and more efficient and safe STING agonist therapy.
Topics: Animals; Mice; Liposomes; Prodrugs; Melanoma; Cell Line, Tumor; Tumor Microenvironment; Immunotherapy
PubMed: 37524727
DOI: 10.1038/s41467-023-40312-y -
Cancer Cell Nov 2023Diffuse intrinsic pontine glioma (DIPG) is an aggressive brain stem tumor and the leading cause of pediatric cancer-related death. To date, these tumors remain...
Diffuse intrinsic pontine glioma (DIPG) is an aggressive brain stem tumor and the leading cause of pediatric cancer-related death. To date, these tumors remain incurable, underscoring the need for efficacious therapies. In this study, we demonstrate that the immune checkpoint TIM-3 (HAVCR2) is highly expressed in both tumor cells and microenvironmental cells, mainly microglia and macrophages, in DIPG. We show that inhibition of TIM-3 in syngeneic models of DIPG prolongs survival and produces long-term survivors free of disease that harbor immune memory. This antitumor effect is driven by the direct effect of TIM-3 inhibition in tumor cells, the coordinated action of several immune cell populations, and the secretion of chemokines/cytokines that create a proinflammatory tumor microenvironment favoring a potent antitumor immune response. This work uncovers TIM-3 as a bona fide target in DIPG and supports its clinical translation.
Topics: Humans; Child; Diffuse Intrinsic Pontine Glioma; Glioma; Immunologic Memory; Hepatitis A Virus Cellular Receptor 2; Brain Stem Neoplasms; Tumor Microenvironment
PubMed: 37802053
DOI: 10.1016/j.ccell.2023.09.001 -
Gut Nov 2023Gut microbiota is a key player in dictating immunotherapy response. We aimed to explore the immunomodulatory effect of probiotic and its role in improving...
OBJECTIVE
Gut microbiota is a key player in dictating immunotherapy response. We aimed to explore the immunomodulatory effect of probiotic and its role in improving anti-programmed cell death protein 1 (PD1) efficacy against colorectal cancer (CRC).
DESIGN
The effects of in anti-PD1 response were assessed in syngeneic mouse models and azoxymethane/dextran sulfate sodium-induced CRC model. The change of immune landscape was identified by multicolour flow cytometry and validated by immunohistochemistry staining and in vitro functional assays. Liquid chromatography-mass spectrometry was performed to identify the functional metabolites.
RESULTS
significantly improved anti-PD1 efficacy in two syngeneic mouse models with different microsatellite instability (MSI) statuses (MSI-high for MC38, MSI-low for CT26). Such effect was confirmed in CRC tumourigenesis model. synergised with anti-PD1 therapy by reducing Foxp3 CD25 regulatory T cell (Treg) intratumoural infiltration, and enhancing effector function of CD8 T cells. -derived indole-3-carboxylic acid (ICA) was identified as the functional metabolite. Mechanistically, ICA inhibited indoleamine 2,3-dioxygenase (IDO1) expression, therefore suppressing kynurenine (Kyn) production in tumours. ICA also competed with Kyn for binding site on aryl hydrocarbon receptor (AHR) and antagonised Kyn binding on CD4 T cells, thereby inhibiting Treg differentiation in vitro. ICA phenocopied effect and significantly improved anti-PD1 efficacy in vivo, which could be reversed by Kyn supplementation.
CONCLUSION
-derived ICA improved anti-PD1 efficacy in CRC through suppressing CD4+Treg differentiation and enhancing CD8+T cell function by modulating the IDO1/Kyn/AHR axis. is a potential adjuvant to augment anti-PD1 efficacy against CRC.
Topics: Animals; Mice; CD8-Positive T-Lymphocytes; Colorectal Neoplasms; Kynurenine; Receptors, Aryl Hydrocarbon; T-Lymphocytes, Regulatory; Lactobacillus; Programmed Cell Death 1 Receptor; Immune Checkpoint Inhibitors; Bacterial Lysates
PubMed: 37770127
DOI: 10.1136/gutjnl-2023-329543 -
Phagocytosis increases an oxidative metabolic and immune suppressive signature in tumor macrophages.The Journal of Experimental Medicine Jun 2023Phagocytosis is a key macrophage function, but how phagocytosis shapes tumor-associated macrophage (TAM) phenotypes and heterogeneity in solid tumors remains unclear....
Phagocytosis is a key macrophage function, but how phagocytosis shapes tumor-associated macrophage (TAM) phenotypes and heterogeneity in solid tumors remains unclear. Here, we utilized both syngeneic and novel autochthonous lung tumor models in which neoplastic cells express the fluorophore tdTomato (tdTom) to identify TAMs that have phagocytosed neoplastic cells in vivo. Phagocytic tdTompos TAMs upregulated antigen presentation and anti-inflammatory proteins, but downregulated classic proinflammatory effectors compared to tdTomneg TAMs. Single-cell transcriptomic profiling identified TAM subset-specific and common gene expression changes associated with phagocytosis. We uncover a phagocytic signature that is predominated by oxidative phosphorylation (OXPHOS), ribosomal, and metabolic genes, and this signature correlates with worse clinical outcome in human lung cancer. Expression of OXPHOS proteins, mitochondrial content, and functional utilization of OXPHOS were increased in tdTompos TAMs. tdTompos tumor dendritic cells also display similar metabolic changes. Our identification of phagocytic TAMs as a distinct myeloid cell state links phagocytosis of neoplastic cells in vivo with OXPHOS and tumor-promoting phenotypes.
Topics: Humans; Macrophages; Phagocytosis; Lung Neoplasms; Myeloid Cells; Oxidative Stress; Tumor Microenvironment
PubMed: 36995340
DOI: 10.1084/jem.20221472 -
Journal of Translational Medicine Sep 2023The chimeric antigen receptor (CAR)-T therapy has a limited therapeutic effect on solid tumors owing to the limited CAR-T cell infiltration into solid tumors and the...
BACKGROUND
The chimeric antigen receptor (CAR)-T therapy has a limited therapeutic effect on solid tumors owing to the limited CAR-T cell infiltration into solid tumors and the inactivation of CAR-T cells by the immunosuppressive tumor microenvironment. Macrophage is an important component of the innate and adaptive immunity, and its unique phagocytic function has been explored to construct CAR macrophages (CAR-Ms) against solid tumors. This study aimed to investigate the therapeutic application of CAR-Ms in ovarian cancer.
METHODS
In this study, we constructed novel CAR structures, which consisted of humanized anti-HER2 or CD47 scFv, CD8 hinge region and transmembrane domains, as well as the 4-1BB and CD3ζ intracellular domains. We examined the phagocytosis of HER2 CAR-M and CD47 CAR-M on ovarian cancer cells and the promotion of adaptive immunity. Two syngeneic tumor models were used to estimate the in vivo antitumor activity of HER2 CAR-M and CD47 CAR-M.
RESULTS
We constructed CAR-Ms targeting HER2 and CD47 and verified their phagocytic ability to ovarian cancer cells in vivo and in vitro. The constructed CAR-Ms showed antigen-specific phagocytosis of ovarian cancer cells in vitro and could activate CD8 cytotoxic T lymphocyte (CTL) to secrete various anti-tumor factors. For the in vivo model, mice with human-like immune systems were used. We found that CAR-Ms enhanced CD8 T cell activation, affected tumor-associated macrophage (TAM) phenotype, and led to tumor regression.
CONCLUSIONS
We demonstrated the inhibition effect of our constructed novel HER2 CAR-M and CD47 CAR-M on target antigen-positive ovarian cancer in vitro and in vivo, and preliminarily verified that this inhibitory effect is due to phagocytosis, promotion of adaptive immunity and effect on tumor microenvironment.
Topics: Humans; Female; Animals; Mice; CD47 Antigen; Ovarian Neoplasms; Macrophages; Phagocytosis; Tumor Microenvironment
PubMed: 37740183
DOI: 10.1186/s12967-023-04479-8