Quantitative Lactylation Modification Proteomics of AC16
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo Sapiens
SUBMITTER: 
Peiqing Liu
PROVIDER: PXD075014 | iProX | Fri Feb 27 00:00:00 GMT 2026
REPOSITORIES: iProX
Similar Datasets
Project description:Nucleus pulposus (NP) resides in hypoxic microenvironment due to the avascular structure of intervertebral disc (IVD). Intracellular lactate drives lysine lactylation (Kla) as a newly epigenetic modification. However, the impact of Kla on NPCs remains unknown. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we showed a global lactylome profiling on NPCs cultured in normoxia and hypoxia environment and found that 3510 lactylation sites on 1052 proteins in NPCs on non-histone proteins. Moreover, there are 18 proteins with 129 Kla sites exclusively detected in the normoxia group, and 117 Kla sites in 27 proteins were specific in hypoxia group. Together, our dates reveals that Kla plays an important role in regulating cellular metabolism and may contribute to IDD progression.
2025-05-06 | PXD048244 | Pride
Project description:Hypoxia promotes tumorigenesis and lactate accumulation in esophageal squamous cell carcinoma (ESCC). Lactate can induce histone lysine lactylation (Kla, a recently-identified histone marks) to regulate transcription. However, the functional consequence of histone Kla under hypoxia in ESCC remains to be explored. Here, we reveal that hypoxia facilitates histone H3K9la to enhance LAMC2 transcription for proliferation of ESCC. We found that global level of Kla was elevated under hypoxia, and thus identified the landscape of histone Kla in ESCC by quantitative proteomics. Furthermore, we show a significant increase of H3K9la level induced by hypoxia. Next, MNseq ChIP-seq and RNA-seq analysis suggest that H3K9la is enriched at the promoter of cell junction genes. Finally, we demonstrate that the histone H3K9la facilitates the expression of LAMC2 for ESCC invasion by in vivo and in vitro experiments. Briefly, our study reveal a vital role of histone Kla triggered by hypoxia in cancer.
2024-05-22 | GSE255352 | GEO
Project description:Cardiovascular diseases are associated with an altered cardiomyocyte metabolism. Due to a shortage of human heart tissue, experimental studies mostly rely on alternative approaches including animal and cell culture models. Since the use of isolated primary cardiomyocytes is limited, immortalized cardiomyocyte cell lines may represent a useful tool as they closely mimic human cardiomyocytes. This study is focused on the AC16 cell line generated from adult human ventricular cardiomyocytes. Despite an increasing number of articles employing AC16 cells, the comprehensive proteomic, bioenergetic and oxygen-sensing characterization of proliferating versus differentiated cells is still lacking. Here, we provide a comparison of these two stages, particularly emphasizing cell metabolism, mitochondrial function, and hypoxic signalling. The label-free quantitative mass spectrometry revealed a decrease in autophagy and cytoplasmic translation in differentiated AC16, confirming their phenotype. Cell differentiation led to the global increase in mitochondrial proteins (e.g. OXPHOS proteins, TFAM, VWA8, etc.) reflected by elevated mitochondrial respiration. Fatty acid oxidation proteins were increased in differentiated cells, while the expression levels of proteins associated with fatty acid synthesis were unchanged, and glycolytic proteins were decreased. There was a profound difference between proliferating and differentiated cells in their response to hypoxia and anoxia/reoxygenation. We conclude that AC16 differentiation leads to proteomic and metabolic shifts and altered cell response to oxygen deprivation. This underscores the requirement for proper selection of particular differentiation state during experimental planning.
2025-04-14 | PXD059985 | Pride
Project description:Emerging evidence suggests that accumulated D-lactate (D-la) induces diverse effects in human intestinal injury diseases. However, the regulatory mechanism remain elusive. Here, we discover the lysine D-lactylation (D-Kla) induced by D-la and reveal the molecular mechanism underlying D-la-driven D-Kla for transcription regulation. We first discovered D-Kla in HepG2 cells stimulated by D-la from E. coli secretions. Next, we validated the existence of D-la-driven D-Kla by four approaches, especially including metabolic labelling experiments. Additionally, we uncover that SCOT1 catalyzes the transformation of D-la to D-lactyl-CoA, serving as a key donor for D-Kla. Furthermore, we identify the D-Kla landscape with 2895 sites by proteomics analysis and 34 histone D-Kla sites activated by D-la and further indicate the regulation of H3K18-Dla in transcription by CUT&Tag and RNA-seq assays. Finally, we show the effect of D-Kla on inhibiting the hepatocellular carcinoma cells proliferation. In summary, we uncover the D-la-driven D-Kla and reveal a D-Kla-mediated transcription regulation for tumor suppression, providing new insight into the protein modifications induced by microbiota secretions.
2026-07-15 | GSE269963 | GEO
Project description:Hypoxia is a hallmark of the tumor microenvironment, but its role in immune evasion in esophageal squamous cell carcinoma (ESCC) remains to be fully elucidated. Here, we demonstrate hypoxia inversely correlated with anti-tumor immune signatures and CD8+ T cell infiltration in clinical samples and murine models. Functionally, reducing hypoxia with the agents TH-302 or PX-478 in the AKR model enhanced intratumoral CD8+ T cell infiltration and increased their expression of Granzyme B, IFNγ, and TNFα. Mechanistically, hypoxia-induced immune suppression was dependent on protein lactylation. Inhibiting lactylation reversed the hypoxic suppression of CD8+ T cell function and abrogated the hypoxia-driven transcriptional program, which involved pathways like glycolysis, TGFβ, and Notch signaling. This lactylation-dependent regulation operated by facilitating the formation of the HIF1 transcription complex. Specifically, mass spectrometry identified lactylation at the K172 site of HIF1α, which was crucial for its binding to HIF1β and subsequent target gene activation. Furthermore, in a preclinical ESCC model, pharmacological inhibition of HIF1α with PX-478 synergized with anti-PD-1 therapy, leading to superior tumor control and enhanced CD8+ T cell cytotoxicity. Our study identifies HIF1α K172 lactylation as a pivotal mechanism of hypoxia-mediated immune escape in ESCC, suggesting a therapeutic strategy to improve immunotherapy.
2026-01-01 | GSE315097 | GEO
Project description:Hypoxia is a hallmark of the tumor microenvironment, but its role in immune evasion in esophageal squamous cell carcinoma (ESCC) remains to be fully elucidated. Here, we demonstrate hypoxia inversely correlated with anti-tumor immune signatures and CD8+ T cell infiltration in clinical samples and murine models. Functionally, reducing hypoxia with the agents TH-302 or PX-478 in the AKR model enhanced intratumoral CD8+ T cell infiltration and increased their expression of Granzyme B, IFNγ, and TNFα. Mechanistically, hypoxia-induced immune suppression was dependent on protein lactylation. Inhibiting lactylation reversed the hypoxic suppression of CD8+ T cell function and abrogated the hypoxia-driven transcriptional program, which involved pathways like glycolysis, TGFβ, and Notch signaling. This lactylation-dependent regulation operated by facilitating the formation of the HIF1 transcription complex. Specifically, mass spectrometry identified lactylation at the K172 site of HIF1α, which was crucial for its binding to HIF1β and subsequent target gene activation. Furthermore, in a preclinical ESCC model, pharmacological inhibition of HIF1α with PX-478 synergized with anti-PD-1 therapy, leading to superior tumor control and enhanced CD8+ T cell cytotoxicity. Our study identifies HIF1α K172 lactylation as a pivotal mechanism of hypoxia-mediated immune escape in ESCC, suggesting a therapeutic strategy to improve immunotherapy.
2026-01-01 | GSE315098 | GEO
Project description:Hypoxia is a hallmark of the tumor microenvironment, but its role in immune evasion in esophageal squamous cell carcinoma (ESCC) remains to be fully elucidated. Here, we demonstrate hypoxia inversely correlated with anti-tumor immune signatures and CD8+ T cell infiltration in clinical samples and murine models. Functionally, reducing hypoxia with the agents TH-302 or PX-478 in the AKR model enhanced intratumoral CD8+ T cell infiltration and increased their expression of Granzyme B, IFNγ, and TNFα. Mechanistically, hypoxia-induced immune suppression was dependent on protein lactylation. Inhibiting lactylation reversed the hypoxic suppression of CD8+ T cell function and abrogated the hypoxia-driven transcriptional program, which involved pathways like glycolysis, TGFβ, and Notch signaling. This lactylation-dependent regulation operated by facilitating the formation of the HIF1 transcription complex. Specifically, mass spectrometry identified lactylation at the K172 site of HIF1α, which was crucial for its binding to HIF1β and subsequent target gene activation. Furthermore, in a preclinical ESCC model, pharmacological inhibition of HIF1α with PX-478 synergized with anti-PD-1 therapy, leading to superior tumor control and enhanced CD8+ T cell cytotoxicity. Our study identifies HIF1α K172 lactylation as a pivotal mechanism of hypoxia-mediated immune escape in ESCC, suggesting a therapeutic strategy to improve immunotherapy.
2026-01-01 | GSE315099 | GEO
Project description:Multiple myeloma (MM) is a hematologic malignancy characterized by abnormal plasma cell proliferation, with lenalidomide emerging as a primary treatment. However, prolonged use often leads to drug resistance, underscoring the need to understand resistance mechanisms. Protein post-translational modifications (PTMs) play crucial roles in disease development, including chemoresistance. Here, we investigate the involvement of new types of PTMs, focusing on lysine lactylation (Kla), in lenalidomide resistance. We observed elevated Kla levels in lenalidomide-resistant MM cells, implicating its role in resistance. Through quantitative proteome, lactylome, and acetylome analysis, we identified 7493 proteins, 1241 Kla sites, and 9313 lysine acetylation (Kac) sites, thereby revealing differential protein expression and PTM profiles in lenalidomide-resistant cells. Proteomic analysis revealed that a serious of chemoresistance related proteins were upregulated, and a number of CRL4-CRBN regulatory factors were downregulated. Lactylome analysis revealed that numerous chemoresistance related proteins exhibited increased Kla levels in lenalidomide-resistant MM cells, suggesting that Kla played an important role in the development of lenalidomide-resistance in LenR MM cells. Notably, histone H4K8la were associated with upregulation of chemoresistance-related genes CDK6 and ECHS1. Our findings shed light on the epigenetic mechanisms underlying lenalidomide resistance in MM, offering insights for overcoming chemoresistance.
2026-04-09 | GSE294374 | GEO
Project description:Multiple myeloma (MM) is a hematologic malignancy characterized by abnormal plasma cell proliferation, with lenalidomide emerging as a primary treatment. However, prolonged use often leads to drug resistance, underscoring the need to understand resistance mechanisms. Protein post-translational modifications (PTMs) play crucial roles in disease development, including chemoresistance. Here, we investigate the involvement of new types of PTMs, focusing on lysine lactylation (Kla), in lenalidomide resistance. We observed elevated Kla levels in lenalidomide-resistant MM cells, implicating its role in resistance. Through quantitative proteome, lactylome, and acetylome analysis, we identified 7493 proteins, 1241 Kla sites, and 9313 lysine acetylation (Kac) sites, thereby revealing differential protein expression and PTM profiles in lenalidomide-resistant cells. Proteomic analysis revealed that a serious of chemoresistance related proteins were upregulated, and a number of CRL4CRBN regulatory factors were downregulated. Lactylome analysis revealed that numerous chemoresistance related proteins exhibited increased Kla levels in lenalidomide-resistant MM cells, suggesting that Kla played an important role in the development of lenalidomide-resistance in LenR MM cells. Notably, histone H4K8la were associated with upregulation of chemoresistance-related genes CDK6 and ECHS1. Our findings shed light on the epigenetic mechanisms underlying lenalidomide resistance in MM, offering insights for overcoming chemoresistance.
2026-04-09 | GSE294373 | GEO
Project description:The formation of cartilaginous organoids fostered by the cell–hydrogel adaptation creates a hypoxic microenvironment and induces metabolic switching to favor glycolysis and the accumulation of lactate in the MSC spheroids, leading to significantly elevated histone lysine lactylation (Kla). We further show that the increased H3K18la enhances chondrogenesis and cartilaginous matrix deposition by regulating the transcriptional accessibility of chondrogenic genes.
2025-03-26 | GSE280956 | GEO