Metabolic dysregulation has been recognized as a crucial driver of tumorigenesis, particularly in metabolic dysfunction-associated steatotic liver disease (MASLD)-related hepatocellular carcinoma (HCC). However, the underlying mechanisms remain poorly understood. Here, we identify lysosome-localized insulin receptor tyrosine kinase substrate (IRTKS) as a key activator of the metabolic master regulator mTORC1 through phospho-antibody array screening. IRTKS forms membrane-associated condensates that selectively interact with the GTPase RRAGD, a key upstream regulator of mTORC1, thereby enhancing the sensitivity of mTORC1 to free amino acids. Notably, in hepatic knockin mice, Irtks-mediated mTORC1 hyperactivation promotes obesity, hepatic lipid accumulation, and the progression from MASLD to metabolic dysfunction-associated steatohepatitis and HCC. Conversely, pharmacological inhibition of mTORC1 or genetic ablation of Irtks ameliorates hepatic steatosis, inflammation, and metabolic dysfunction in mouse models. Our study establishes IRTKS as a central regulator of mTORC1-dependent metabolic reprogramming during hepatocarcinogenesis, providing potential therapeutic targets for MASLD-associated liver cancer.

Abstract The overall incidence of extramedullary disease (EMD) in acute myeloid leukemia (AML) ranges from 2% to 30.5%, and is potentially associated with short overall survival. However, the risk factors for EMD and the underlying molecular mechanisms remain poorly understood. Here, we analyzed a cohort of 118 adult patients with de novo AML, among whom 27 (22.88%) developed EMD in various body sites, with tissue involvement as the most common (16/118 [13.56%]). Mesothelin (MSLN) expression, detectable at both transcript and protein levels, can serve as an independent risk factor for EMD in patients with AML. Multivariate analyses revealed that MSLN positivity at transcript levels (hazard ratio [HR], 3.43; 95% confidence interval [CI], 1.44-8.20; P = .006) and MSLN positivity on leukemic blasts detected by flow cytometry (HR, 3.28; 95% CI, 1.37-7.86; P = .008) are independently correlated with EMD risk. Mechanistically, MSLN overexpression promotes cell proliferation, metastasis, and invasion in AML cells. MSLN greatly facilitates cell-cell adhesion by binding with mucin 16 (MUC16), resulting in extramedullary dissemination. MSLN also activates the phosphoinositide 3-kinase (PI3K) signaling pathway and upregulates neural cell adhesion molecules (CAMs) CD56 and neural cell adhesion molecule 2 (NCAM2) through interaction with MUC16. Our study highlights the importance of MSLN expression, particularly identified by flow cytometry, as an independent risk factor for EMD in patients with AML, and elucidates its role in modulating PI3K signaling and CAMs in AML cells.

ARID1A is frequently mutated in non-tumorous human tissues and various cancers. However, its role in metabolic dysfunction-associated steatohepatitis (MASH) and hepatocellular carcinoma (HCC) onset and progression remains controversial. Given the critical role of the gut-liver axis in MASH and HCC, we hypothesized that the gut microbiota might contribute to ARID1A deficiency-induced liver diseases. Here, we demonstrate that liver-specific Arid1a deficiency drives MASH/HCC development through dysregulated bile acid metabolism and gut microbiota restructuring. Gut microbiota depletion using an antibiotic cocktail attenuates MASH progression and reduces HCC incidence in liver-specific Arid1a-deficient mice. While lipopolysaccharide (LPS) accumulation due to gut barrier dysfunction promotes HCC progression, it does not significantly affect MASH activity or tumor initiation in Arid1a-deficient mice. Mechanistically, ARID1A deficiency reduces chromatin accessibility and impairs SWI/SNF complex binding at the promoter of nuclear receptor FXR, which governs bile acid homeostasis, suppressing FXR transcription. Further analyses in Arid1a-deficient mice reveal altered gut microbiota composition enriched in bile salt hydrolase (BSH) genes and bile acid profiles showing elevated levels of the secondary bile acid taurodeoxycholic acid (TDCA). When administered to mice, TDCA exacerbates liver inflammation and fibrosis specifically in Arid1a-deficient mice by promoting neutrophil infiltration and hepatic stellate cell activation. Therapeutic interventions including vancomycin to eliminate bile acid-metabolizing bacteria, obeticholic acid to activate Fxr, and cholestyramine to excrete bile acids substantially ameliorate MASH pathology in Arid1a-deficient mice. Collectively, our findings establish the ARID1A-FXR-bile acid-gut microbiota axis as a pathogenic driver of MASH and HCC, offering mechanism-based therapeutic strategies for patients with ARID1A mutations.

The mechanisms that regulate immune evasion by pancreatic ductal adenocarcinomas (PDACs) remain poorly understood. Using a mouse model of resectable PDAC, we identified an unknown role of the circadian rhythm gene Differentially Expressed in Chondrocytes 2 (Dec2) in regulating tumor progression and dormancy. Deletion of Dec2 from tumor cells substantially increased mouse survival after resection due to an immune-mediated mechanism, as the survival benefit was abrogated under immunodeficient conditions. Dec2 promotes immune evasion by repressing major histocompatibility complex class I (MHC-I)-dependent antigen presentation and by repolarizing the tumor microenvironment from immunologically cold (low T cell infiltration) to hot (elevated T cell infiltration). Dec2 is also a regulator of circadian rhythms, and we found that genes involved in MHC-I antigen presentation and MHC-I surface localization oscillated in a circadian manner, which was lost upon deletion of Dec2 in vitro. We conclude that Dec2 promotes primary PDAC progression and likely metastatic dormancy through immune evasion.

C-type lectin-like receptor 2 (CLEC2) is a transmembrane receptor highly expressed on platelets, which regulates platelet aggregation and immune response. Yet, the function of CLEC2 in lung epithelium and its contribution to acute lung injury (ALI) is unclear. In this study, a lung epithelial-specific CLEC2 knockout mouse (Clec1bAT2-KO) was generated and performed for ALI models. In both LPS- and acid-induced lung injury models, the ALI signs of Clec1bAT2-KO mice were further exacerbated. The therapeutic application of epithelial-restricted CLEC2 overexpression using adeno-associated virus (AAV) or CLEC2 activation using its endogenous ligand podoplanin serves as a lung epithelial protective agent in the setting of ALI. Transcriptomic analyses reveal that CLEC2-regulated genes are highly enriched in chemotaxis, cytokine, and extracellular matrix (ECM) components. Lung injury was partially attenuated in Ccl5-/-, Csf3-/-, and Cxcl1-/- mice pretreated with AAV-si-CLEC2, followed by LPS challenge. Loss of CLEC2 leads to ECM degradation, which could be reversed by exogenous transforming growth factor beta (TGF-β). Furthermore, interferon regulatory factor 1 (IRF1) was identified as the key molecule that regulates CLEC2-related cytokine/chemokine production and ECM degradation. These findings suggest that epithelial CLEC2 protects against ALI by modulating spleen tyrosine kinase/IRF1-mediated cytokine/chemokine production and TGF-β-mediated ECM remodeling.

SCARA5 (Scavenger Receptor Class A Member 5), a member of scavenger receptor class A, is a type II transmembrane protein. Previous studies, including our own, have suggested that SCARA5 acts as a tumor suppressor in various cancers. Additionally, SCARA5 has been identified as a ferritin receptor that facilitates iron delivery independent of transferrin. However, it remains unclear whether ferroptosis is involved in the tumor-suppressive function of SCARA5 in hepatocellular carcinoma (HCC). In this study, we found that SCARA5-deficient cells, including mouse embryonic fibroblasts (MEFs) and HCC cells, exhibited reduced sensitivity to ferroptosis induced by erastin and RSL3. We measured the cell viability, cellular reactive oxygen species (ROS), lipid ROS, malondialdehyde (MDA) and ferrous iron concentration to assess the role of SCARA5 in ferroptosis. Mechanistically, we confirmed that SCARA5 might enhance the intracellular availability of bioactive ferrous iron by promoting autophagic degradation of the major iron storage protein ferritin. Furthermore, we found that SCARA5 deficiency contributed to the resistance of HCC cells to sorafenib, a therapeutic agent for HCC, possibly by inhibiting ferroptosis. Collectively, our study revealed the role of SCARA5 in regulating ferroptosis, providing a profound understanding of sorafenib resistance in HCC systemic therapy.

The mechanisms that regulate cancer dormancy remain poorly understood. Using an orthotopic mouse model of resectable pancreatic adenocarcinoma (PDAC), we identified the transcriptional repressor factor Dec2 (Bhlhe41) as a gene that was upregulated in metastatic dormant tumor cells from the liver. To understand how Dec2 regulates dormancy, we deleted it from the murine pancreatic cancer cells and implanted them in the model. Following resection, mice implanted with Dec2 KO cells demonstrated a striking improvement in survival compared to the mice with Dec2 WT cells. This difference in survival was abrogated when nude mice were used as hosts indicating the survival difference is from an immune mediated mechanism. Pancreatic cancer is a highly immunosuppressive tumor due to both a relatively low expression of major histocompatibility complex class I (MHC-I)-dependent antigen presentation and an abundance of myeloid-derived suppressor cells that lead to poor T cell trafficking and activation. We found that deletion of Dec2 overcame both of these mechanisms to restore anti-tumor immunity that inhibited tumor growth not only in primary tumors as well as metastases. Dec2 suppresses MHC-I antigen presentation by repressing multiple genes in the antigen presentation pathway including H2-k1, H2-d1, Tap1, and B2m as well as the source of tumor antigens from the proteosome by repressing multiple proteasome genes. Loss of Dec2 repolarized the tumor immune microenvironment by decreasing expression of multiple immunosuppressive cytokines such as Cxcl1, Ccl2 and Csf2 in tumors. We conclude that Dec2 facilitates both pancreatic tumor growth and dormancy by regulating tumor cell MHCI antigen presentation. Dec2 is known to function as part of a negative accessory arm of the molecular circadian clock. We found several components of the antigen presentation pathway oscillated in a circadian manner that was lost upon deletion of Dec2. Moreover, T-cell mediated tumor cell killing varied depending on the time of day. We suggest that lowered MHC-I presentation of antigens during rest phase is a natural effect of the circadian clock. Lan Wang, Chris Harris, Orjola Prela, Wade Narrow, Jennifer L. Becker, Juliana Cazarin de Menezes, Darren R. Carpizo. The circadian gene Dec2 promotes pancreatic cancer dormancy by regulating tumor cell antigen presentation to facilitate immune evasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 4048.

As a predominant oncogenic driver in non-small cell lung cancer (NSCLC), EGFR frequently undergoes amplification or mutation, with EGFR-tyrosine kinase inhibitors (EGFR-TKIs) like gefitinib and erlotinib constituting frontline therapy for advanced EGFR-mutant cases. However, both primary and acquired resistance significantly limit clinical efficacy. Here, we revealed that glutathione metabolic pathway controlled by glutathione peroxidase GPX2 was abnormally activated in gefitinib-resistant A549 and HCC827-GR cell lines. Mechanistically, GPX2 triggers Hedgehog signaling activation through releasing GLI transcriptional regulator, promoting cancer stem cell (CSC) characteristics and TKI resistance. Notably, N6-methyladenosine (m6A) modification on GPX2 mRNA mediated by METTL14 diminished its stability. In vivo, GPX2 deletion constrained glutathione metabolism and boosted the effectiveness of TKI in cell line-derived xenograft models. Collectively, these findings demonstrate that GPX2 serves as a positive regulator of both primary and acquired EGFR-TKI resistance and could be a promising therapeutic target for precise treatment of NSCLC.

5-Fluorouracil (5-FU) is the primary chemotherapeutic agent for the clinical management of advanced gastric cancer (GC). However, the emergence of drug resistance remains an inescapable challenge. In drug-resistant cancer cells, prior treatments contribute to elevated oxidative stress, resulting in higher level of reactive oxygen species (ROS) compared to treatment-naïve cancer cells. Activation of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) has been shown to promote ROS production and facilitate epithelial-mesenchymal transition in GC. In this study, we found that LOX-1 silencing significantly increased 5-FU sensitivity by reducing tumor cell viability and colony-forming ability. Enrichment analysis suggested that nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) might act as a downstream effector of LOX-1, and overexpression of NOX4 was able to counteract the increased sensitivity to 5-FU induced by LOX-1 depletion. Additionally, bioinformatic predictions and in vitro experiments indicated that LOX-1 regulated NOX4 expression through JAK2/STAT3 signaling pathway. Altogether, this study provides novel evidence that LOX-1 mediated JAK2/STAT3/NOX4 axis plays a crucial role in modulating 5-FU sensitivity of GC and targeting LOX-1 may offer a promising therapeutic strategy to enhance the efficacy of 5-FU in advanced gastric cancer treatment.

ARID1A, which encodes an important subunit of SWI/SNF complex, is frequently mutated in non-malignant tissues and tumors. However, how ARID1A loss enables environmental carcinogens to initiate tumorigenesis remains unknown. Here, liver-specific Arid1a-deficient (Arid1aLKO) mice are exposed to aristolochic acid I (AAI), a potent herbal carcinogen. Notably, AAI dramatically accelerated hepatocarcinogenesis in Arid1a-deficient livers, accompanied by a specific 3' splice-site mutation in Ctnnb1 in most tumors and adjacent non-tumorous tissues. This mutation results in exon 3 skipping and subsequent β-catenin activation. Single-nucleus RNA-seq coupled with phylogenetic analyses reveals AAI-induced tumor microenvironment alteration and clonal expansion of β-catenin-activated cells. Conversely, inhibition of β-catenin signaling significantly suppresses AAI-induced tumors in the context of Arid1a loss. Mechanistically, Arid1a deficiency transcriptionally represses the expression of critical genes related to nucleotide excision repair, which removes AAI-derived DNA adducts, due to SWI/SNF complex dysfunction. Simultaneously, it upregulates Nqo1, a key enzyme enhancing AAI bioactivation and AAI-DNA adduct formation. This dual-hit mechanism, characterized by impaired DNA repair and heightened genotoxicity, explains synergistic carcinogenesis. The study unveils ARID1A as a guardian against environmental carcinogens and proposes β-catenin blockade for precision prevention in high-risk patients with ARID1A-mutant benign liver diseases.

The mechanisms that regulate tumor progression and dormancy in pancreatic adenocarcinoma (PDAC) remain poorly understood. We established a murine model of early-stage PDAC that recapitulates the overall survival and metastatic recurrence patterns of patients. We isolated disseminated tumor cells (DTCs) from the livers of latent recurrent mice and found them with characteristics of dormant cells. Transcriptomic analysis indicated that not only were these cells quiescent (Ki67lo) relative to primary tumor cells, but they also exhibited high levels of the transcriptional repressor Dec2. We validated this finding in human PDAC on liver tissues from patients with stage IV disease demonstrating that isolated small clusters of DTCs were more likely Dec2hi/Ki67lo, than tumor cells in established metastases. Compared to normal pancreas, Dec2 levels are increased in PDAC tumor, and we could detect Dec2 in proliferating PDAC cells (though not at the levels as seen in dormant DTC), suggesting that Dec2 plays a role in PDAC biology beyond dormancy. We generated Dec2 knock out (KO) in murine PDAC cells for orthotopic and liver metastasis models. Loss of Dec2 markedly inhibited tumor progression and metastasis that was dependent on adaptive immune system, as the effect was abrogated when either nude hosts were used, or immunocompetent hosts were subjected to anti-CD4 and CD8 depletion. We further found that Dec2 KO cells had increased surface MHC-I as well as total MHC-I. CUT&RUN indicates that Dec2 transcriptional represses multiples genes involved in antigen presentation including H2-k1, H2-d1, B2M, Tap1 and PSMB 8,9 and 10. We found that proteosome activity was significantly increased in Dec2 KO cells and the inhibition of proteasome activity with MG132 prevented surface MHC-I display in Dec2 KO cells. We also examined the PDAC microenvironment for alterations in immune cell populations using flow cytometry and cytokines using Luminex array. Compared to WT, Dec2 KO tumors had significantly more infiltrating CD8 T cells but fewer monocytes and macrophages, particularly immunosuppressive Arg1+ macrophages. Examination of intratumoral cytokine levels showed that, relative to WT tumors, Dec2 KO had a significant decrease of Cxcl1, Ccl2 and Csf2 that are known to drive PDAC immunosuppression and a significant increase in IFN-γ. These data indicate that the anti-tumor immune mechanism induced by Dec2 KO involves not only regulation of MHC-I but also repolarization of the TME from suppressive to reactive. Taken together, we found that Dec2 plays a unique role in regulating PDAC tumor progression and dormancy. At constitutive levels Dec2 promotes pancreatic cancer progression through creating an immunosuppressive TME and immune evasion, whereas very high level of Dec2 promotes cellular dormancy. Our data identify Dec2 as a novel driver of PDAC tumor progression and dormancy and suggest that targeting Dec2 would have therapeutic potential. Lan Wang, Chris Harris, Orjola Prela, Wade C. Narrow, Brian Habb, Darren Carpizo. Dec2 drives tumor progression by facilitating immune evasion in pancreatic cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research—Emerging Science Driving Transformative Solutions; Boston, MA; 2025 Sep 28-Oct 1; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2025;85(18_Suppl_3):Abstract nr A014.

Alanine Serine Cysteine transporter 2 (ASCT2/SLC1A5) is a key glutamine transporter in cancer cells and has been shown in a variety of cancers to promote tumor growth by reprogramming glutamine metabolism and altering the tumor microenvironment. However, the role in thyroid cancer remains unknown. To investigate the expression and prognostic value of SLC1A5 in thyroid cancer using publically available databases, and to define the relationship with clinical characteristics. SLC1A5 expression in TPC-1 and B-CPAP was knocked down using SLC1A5 siRNA to investigate its effects on cell growth and apoptosis. Transcriptome sequencing and metabolite analysis were carried out in the SLC1A5 siRNA group to identify major transcriptomic or metabolite changes that could lead to apoptosis. In addition, we explored for the connection of SLC1A5 with the tumor microenvironment using algorithms like ESTIMATE and CIBERSORT. High SLC1A5 expression in THCA is related with a poor prognosis and advanced clinical stage. In vitro findings showed that SLC1A5 knockdown reduced THCA cell activity and accelerated apoptosis, and the results were consistent with the effect of SLC1A5 inhibitor GPNA. While RNA sequencing analysis revealed that NF-κb signaling was enhanced and oxidative phosphorylation levels were lowered. Metabolomics findings indicated that Glutathione and purine metabolism were dramatically affected in the SLC1A5 siRNA group. Furthermore, immune microenvironment study revealed that SLC1A5 had a positive correlation with the amount of CD4 + T memory-activated cells and T cell follicular helper cells. SLC1A5 may be a possible target in THCA. Our findings indicate that DEGs and differential metabolites are mostly linked to numerous signaling pathways and immunological modulation, which may play an important role in SLC1A5 regulation of THCA development.

Extracellular vesicles (EVs) are secreted, cell-derived, membrane-bound compartments implicated in various diseases for their ability to influence distant targets and as carriers of biomarkers. Here, we present a protocol for separating EVs from mammalian pancreatic cancer cells and their characterization using western blot and electron microscopy. We then demonstrate how they are utilized to affect tumor development in a murine model of metastatic pancreatic cancer including a method to quantify hepatic tumor burden in histologic samples.For complete details on the use and execution of this protocol, please refer to Dudgeon et al.1

Hepatocellular carcinoma (HCC) is the major form of liver malignancy with high incidence and mortality. Identifying novel biomarkers and understanding regulatory mechanisms underlying the development and progression of HCC are critical for improving diagnosis, treatment and patient outcomes. Carboxyl terminus of Hsc-70-interacting protein (CHIP) is a well-described U-box-type E3 ubiquitin ligase which promotes the ubiquitination and degradation of numerous tumor-associated proteins. Recent studies have shown that CHIP can play as a tumor-suppressor gene or an oncogene in different kinds of malignancies. To date, the function and mechanism of CHIP in hepatocellular carcinoma remains largely unknown. Based on TCGA data, we found that compared with high CHIP expression, the overall survival of HCC patients with low expression of CHIP was better. In addition, CHIP overexpression markedly enhanced HCC cell proliferation and colony formation. Conversely, knockdown of CHIP restrained the proliferation and colony formation of HCC cells. Meanwhile, knockdown of CHIP decreased mitochondrial cristae or ruptured outer mitochondrial membrane, promoted the accumulation of Fe2+ and ferroptosis of HCC cells. Further research for the first time confirmed that CHIP interacts and degrades transferrin receptor 1 (TfR1) by ubiquitin-proteasome pathway, which leads to the inhibition of ferroptosis and promotes the proliferation of HCC cells. The analysis of proteomics data from CPTAC revealed a negative correlation between CHIP and TfR1 protein expression levels in HCC. These findings indicate that CHIP acts as a negative modulator of ferroptosis and functions as an oncogene in HCC.

Background Apolipoprotein B mRNA editing enzyme catalytic polypeptide-like 2 (APOBEC2) is associated with nucleotide alterations in the transcripts of tumor-related genes which are contributed to carcinogenesis. Expression and prognosis value of APOBEC2 in stomach adenocarcinoma (STAD) remains unclear. Methods The APOBEC2 gene alteration frequency of STAD and APOBEC2 gene expression in STAD and normal tissues were investigated in cBioportal and GEPIA, respectively. We detected expression of APOBEC2, infiltration of CD66b + tumor-associated neutrophils and CD163 + tumor-associated macrophages in tissue microarrays by immunohistochemistry. APOBEC2 gene expression was explored by western blot and qRT-PCR. Relationships between APOBEC2 and CD66b, CD163, and other clinicopathological characteristics were investigated. Associations among APOBEC2 expression status and patient survival outcome were further analyzed. Results APOBEC2 gene alteration frequency was 5%, and APOBEC2 gene was downexpressed in STAD compared to normal tissues ( P < 0.05). APOBEC2 expression status were associated with the infiltration of CD66b + TANs, differentiation grade, TNM stage, histological type and gender (all P < 0.05) in STAD. Little or no APOBEC2 expression was detected in STAD and adjacent normal tissues by western blot. We failed to show that APOBEC2 was an independent risk factor for OS (Hazard Ratio 0.816, 95%CI 0.574–1.161, P = 0.259) or DFS (Hazard Ratio 0.821, 95%CI 0.578–1.166, P = 0.270) in STAD by multivariate Cox regression analysis, but APOBEC2 negative subgroup has a worse OS and DFS among patients with adjuvant chemotherapy. Conclusions APOBEC2 correlates with CD66b, differentiation grade, TNM stages, histological classification, and gender in STAD. APOBEC2 is not an independent prognostic factor for STAD, our results suggest that patients with positive APOBEC2 can benefit from postoperative chemotherapy, and combination of APOBEC2 and CD66b is helpful to further stratify patients into different groups with distinct prognoses.

Metabolic reprogramming is one of the essential features of tumors that may dramatically contribute to cancer metastasis. Employing liquid chromatography-tandem mass spectrometry-based metabolomics, we analyzed the metabolic profile from 12 pairwise serum samples of NSCLC brain metastasis patients before and after CyberKnife Stereotactic Radiotherapy. We evaluated the histopathological architecture of 144 surgically resected NSCLC brain metastases. Differential metabolites were screened and conducted for functional clustering and annotation. Metabolomic profiling identified a pathway that was enriched in the metabolism of branched-chain amino acids (BCAAs). Pathologically, adenocarcinoma with a solid growth pattern has a higher propensity for brain metastasis. Patients with high BCAT1 protein levels in lung adenocarcinoma tissues were associated with a poor prognosis. We found that brain NSCLC cells had elevated catabolism of BCAAs, which led to a depletion of α-KG. This depletion, in turn, reduced the expression and activity of the m6A demethylase ALKBH5. Thus, ALKBH5 inhibition participated in maintaining the m6A methylation of mesenchymal genes and promoted the occurrence of epithelial-mesenchymal transition (EMT) in NSCLC cells and the proliferation of NSCLC cells in the brain. BCAA catabolism plays an essential role in the metastasis of NSCLC cells.

High myopia (HM) is a leading cause of blindness worldwide with currently no effective interventions available. A major hurdle lies in its often isolated perception as a purely ocular morbidity, disregarding potential systemic implications. Recent evidence suggests the existence of a gut-eye axis; however, the role of gut microbiota in the pathogenesis of HM remains largely unexplored. Herein, we provide a potential crosstalk among HM's gut dysbiosis, microbial metabolites, and scleral remodeling. Utilizing 16S rRNA gene sequencing, we observed an altered gut microbiota profile in HM patients with a significant reduction in probiotic abundance compared with healthy controls. Subsequent targeted metabolic profiling revealed a notable decrease in plasma levels of the gut microbiota-derived metabolite indole-3-acetic acid (3-IAA) among HM patients, which is closely associated with the reduced probiotics, both negatively correlated with HM severity. Genetic analyses determined that gut microbiota are causally associated with myopia risk. Importantly, when mice subjected to HM modeling receive fecal microbiota transplantation from healthy donors, there is an increase in 3-IAA plasma levels and simultaneous retardation of HM progression along with better maintenance of collagen type I alpha 1 (COL1A1) expression in the sclera. Furthermore, 3-IAA gavage achieves similar effects. Mechanistic investigations confirm the transcriptional activation of COL1A1 by 3-IAA via promoting the enrichment of SP1 to its promoter. Together, our findings provide novel insights into the gut microbiota-eye axis in the pathogenesis of HM and propose new strategies for HM intervention by remodeling the gut microbiota and indole supplementation.