Active Macropinocytosis, Lipid Catabolism, and Exhausting Immune Microenvironment of Ascites Tumor Cells Are Involved in Resistance to Platinum-Based Therapy in Patients With High-Grade Serous Ovarian Cancer

Abstract

Platinum resistance remains a clinical challenge in ovarian cancer. Ascites represents an important mediator and a unique tumor microenvironment (TME) for invasion and metastasis. This study performed high-resolution mass spectrometry (MS) on pre-chemotherapy ascites cells from ovarian cancer patients. Integrating proteomic profiling, clinical data, and single-cell analysis revealed that platinum-resistant ascites displayed a distinct microenvironmental: the macropinocytosis-related protein Src homology 3 domain-containing YSC84-like 1 (SH3YL1) was upregulated, whereas the immune-activation marker CD44 was downregulated in resistant cases. Single-cell analyses and pathway enrichment indicated immune exhaustion in resistant ascites, alongside enhanced macropinocytosis and lipid catabolism in tumor cells. Clinical data also showed that resistant ascites are lipid-rich, with immunofluorescence plus flow cytometry confirming its association with immune exhaustion. Cellular experiments confirmed that SH3YL1-mediated macropinocytosis promoted lipid uptake, and its inhibition partially restored cisplatin sensitivity. A combined model of immune exhaustion, macropinocytosis, and lipid catabolism suggests these ascites-associated features could somewhat predict the platinum sensitivity in ovarian cancer tissues. We therefore propose the hypothesis that, in a lipid-rich ascites microenvironment, immune exhaustion occurs while tumor cells activate macropinocytosis and lipid catabolism—forming a network of resistance mechanisms that may serve as potential predictive markers or intervention targets for platinum resistance.

Keywords

immune microenvironment / lipid metabolism / macropinocytosis / ovarian cancer / platinum resistance

References

[1] B. T. Hennessy, R. L. Coleman, and M. Markman, “Ovarian Cancer,” Lancet 374, no. 9698 (2009): 1371–1382.

[2] H. Sung, J. Ferlay, R. L. Siegel, et al., “Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries,” CA: A Cancer Journal for Clinicians 71, no. 3 (2021): 209–249.

[3] B. van Zyl, D. Tang, and N. A. Bowden, “Biomarkers of Platinum Resistance in Ovarian Cancer: What Can We Use to Improve Treatment,” Endocrine-Related Cancer 25, no. 5 (2018): R303–R318.

[4] A. da Costa and G. Baiocchi, “Genomic Profiling of Platinum-Resistant Ovarian Cancer: The Road Into Druggable Targets,” Seminars in Cancer Biology 77 (2021): 29–41.

[5] M. A. Khan, K. S. Vikramdeo, S. K. Sudan, et al., “Platinum-Resistant Ovarian Cancer: From Drug Resistance Mechanisms to Liquid Biopsy-Based Biomarkers for Disease Management,” Seminars in Cancer Biology 77 (2021): 99–109.

[6] M. Wang, J. Zhang, and Y. Wu, “Tumor Metabolism Rewiring in Epithelial Ovarian Cancer,” Journal of Ovarian Research 16, no. 1 (2023): 108.

[7] L. M. Poisson, A. Munkarah, H. Madi, et al., “A Metabolomic Approach to Identifying Platinum Resistance in Ovarian Cancer,” Journal of Ovarian Research 8 (2015): 13.

[8] S. Alonezi, J. Tusiimire, J. Wallace, et al., “Metabolomic Profiling of the Effects of Melittin on Cisplatin Resistant and Cisplatin Sensitive Ovarian Cancer Cells Using Mass Spectrometry and Biolog Microarray Technology,” Metabolites 6, no. 4 (2016): 35.

[9] T. Worzfeld, E. Pogge von Strandmann, M. Huber, et al., “The Unique Molecular and Cellular Microenvironment of Ovarian Cancer,” Frontiers in Oncology 7 (2017): 24.

[10] K. M. Nieman, H. A. Kenny, C. V. Penicka, et al., “Adipocytes Promote Ovarian Cancer Metastasis and Provide Energy for Rapid Tumor Growth,” Nature Medicine 17, no. 11 (2011): 1498–1503.

[11] A. Bamias, M. L. Tsiatas, E. Kafantari, et al., “Significant Differences of Lymphocytes Isolated From Ascites of Patients With Ovarian Cancer Compared to Blood and Tumor Lymphocytes. Association of CD3+CD56+ Cells With Platinum Resistance,” Gynecologic Oncology 106, no. 1 (2007): 75–81.

[12] L. Zennaro, L. Nicolè, P. Vanzani, F. Cappello, and A. Fassina, “1H-NMR Spectroscopy Metabonomics of Reactive, Ovarian Carcinoma and Hepatocellular Carcinoma Ascites,” Pleura Peritoneum 5, no. 2 (2020): 20200113.

[13] E. Brencicova, A. L. Jagger, H. G. Evans, et al., “Interleukin-10 and Prostaglandin E2 Have Complementary but Distinct Suppressive Effects on Toll-Like Receptor-Mediated Dendritic Cell Activation in Ovarian Carcinoma,” PLoS ONE 12, no. 4 (2017): e0175712.

[14] H.-R. Jin, J. Wang, Z.-J. Wang, et al., “Lipid Metabolic Reprogramming in Tumor Microenvironment: From Mechanisms to Therapeutics,” Journal of Hematology & Oncology 16, no. 1 (2023): 103.

[15] Y. Gong, J. Yang, Y. Wang, L. Xue, and J. Wang, “Metabolic Factors Contribute to T-Cell Inhibition in the Ovarian Cancer Ascites,” International Journal of Cancer 147, no. 7 (2020): 1768–1777.

[16] H. Ponta, L. Sherman, and P. A. Herrlich, “CD44: From Adhesion Molecules to Signalling Regulators,” Nature Reviews Molecular Cell Biology 4, no. 1 (2003): 33–45.

[17] X. Ma, E. Bi, Y. Lu, et al., “Cholesterol Induces CD8+ T Cell Exhaustion in the Tumor Microenvironment,” Cell Metabolism 30, no. 1 (2019): 143–156.e5.

[18] P. Sansone and J. Bromberg, “Targeting the Interleukin-6/Jak/Stat Pathway in Human Malignancies,” Journal of Clinical Oncology 30, no. 9 (2012): 1005–1014.

[19] J. Hasegawa, E. Tokuda, T. Tenno, et al., “SH3YL1 Regulates Dorsal Ruffle Formation by a Novel Phosphoinositide-Binding Domain,” Journal of Cell Biology 193, no. 5 (2011): 901–916.

[20] K. Bouzakri, R. Austin, A. Rune, et al., “Malonyl CoenzymeA Decarboxylase Regulates Lipid and Glucose Metabolism in Human Skeletal Muscle,” Diabetes 57, no. 6 (2008): 1508–1516.

[21] I. Mellman and Y. Yarden, “Endocytosis and Cancer,” Cold Spring Harbor Perspectives in Biology 5, no. 12 (2013): a016949.

[22] J. D. Orth and M. A. McNiven, “Get off My Back! Rapid Receptor Internalization Through Circular Dorsal Ruffles,” Cancer Research 66, no. 23 (2006): 11094–11096.

[23] J. A. Swanson and C. Watts, “Macropinocytosis,” Trends in Cell Biology 5, no. 11 (1995): 424–428.

[24] Z. Derdak, K. A. Villegas, R. Harb, A. M. Wu, A. Sousa, and J. R. Wands, “Inhibition of p53 Attenuates Steatosis and Liver Injury in a Mouse Model of Non-Alcoholic Fatty Liver Disease,” Journal of Hepatology 58, no. 4 (2013): 785–791.

[25] A. N. Urbanek, R. Chan, and K. R. Ayscough, “Function and Interactions of the Ysc84/SH3yl1 Family of Actin- and Lipid-Binding Proteins,” Biochemical Society Transactions 43, no. 1 (2015): 111–116.

[26] E. W. Krueger, J. D. Orth, H. Cao, and M. A. McNiven, “A Dynamin-Cortactin-Arp2/3 Complex Mediates Actin Reorganization in Growth Factor-Stimulated Cells,” Molecular Biology of the Cell 14, no. 3 (2003): 1085–1096.

[27] B. Banushi, S. R. Joseph, B. Lum, J. J. Lee, and F. Simpson, “Endocytosis in Cancer and Cancer Therapy,” Nature Reviews Cancer 23, no. 7 (2023): 450–473.

[28] N. Alaridah, N. Lutay, E. Tenland, et al., “Mycobacteria Manipulate G-Protein-Coupled Receptors to Increase Mucosal Rac1 Expression in the Lungs,” Journal of Innate Immunity 9, no. 3 (2017): 318–329.

[29] S. Mylvaganam, S. A. Freeman, and S. Grinstein, “The Cytoskeleton in Phagocytosis and Macropinocytosis,” Current Biology 31, no. 10 (2021): R619–R632.

[30] M. Kobayashi, K. Harada, M. Negishi, and H. Katoh, “Dock4 Forms a Complex With SH3YL1 and Regulates Cancer Cell Migration,” Cellular Signalling 26, no. 5 (2014): 1082–1088.

[31] E. Stur, S. Corvigno, M. Xu, et al., “Spatially Resolved Transcriptomics of High-Grade Serous Ovarian Carcinoma,” iScience 25, no. 3 (2022): 103923.

[32] D. Trivanović, S. Vignjević Petrinović, I. Okić Djordjević, T. Kukolj, D. Bugarski, and A. Jauković, “Adipogenesis in Different Body Depots and Tumor Development,” Frontiers in Cell and Developmental Biology 8 (2020): 571648.

[33] D. Ishay-Ronen, M. Diepenbruck, R. K. R. Kalathur, et al., “Gain Fat-Lose Metastasis: Converting Invasive Breast Cancer Cells Into Adipocytes Inhibits Cancer Metastasis,” Cancer Cell 35, no. 1 (2019): 17–32.e6.

[34] X. Zheng, X. Wang, X. Cheng, et al., “Single-Cell Analyses Implicate Ascites in Remodeling the Ecosystems of Primary and Metastatic Tumors in Ovarian Cancer,” Nature Cancer 4 (2023): 1138–1156.

[35] L. Zheng, S. Qin, W. Si, et al., “Pan-Cancer Single-Cell Landscape of Tumor-Infiltrating T Cells,” Science 374, no. 6574 (2021): abe6474.

[36] Z. Liu, Z. Yang, J. Wu, et al., “A Single-Cell Atlas Reveals Immune Heterogeneity in Anti-PD-1-Treated Non-Small Cell Lung Cancer,” Cell 188, no. 11 (2025): 3081–3096.e19.

[37] Y. Yao, X.-H. Xu, and L. Jin, “Macrophage Polarization in Physiological and Pathological Pregnancy,” Frontiers in Immunology 10 (2019): 792.

[38] K. J. Warren, D. Iwami, D. G. Harris, J. S. Bromberg, and B. E. Burrell, “Laminins Affect T Cell Trafficking and Allograft Fate,” Journal of Clinical Investigation 124, no. 5 (2014): 2204–2218.

[39] D. Avtanski, K. Chen, and L. Poretsky, “Resistin and Adenylyl Cyclase-Associated Protein 1 (CAP1) Regulate the Expression of Genes Related to Insulin Resistance in BNL CL.2 Mouse Liver Cells,” Data Brief 25 (2019): 104112.

[40] B. G. Dorner, M. B. Dorner, X. Zhou, et al., “Selective Expression of the Chemokine Receptor XCR1 on Cross-Presenting Dendritic Cells Determines Cooperation With CD8+ T Cells,” Immunity 31, no. 5 (2009): 823–833.

[41] H. Karsunky, M. Merad, A. Cozzio, I. L. Weissman, and M. G. Manz, “Flt3 Ligand Regulates Dendritic Cell Development From Flt3+ Lymphoid and Myeloid-Committed Progenitors to Flt3+ Dendritic Cells In Vivo,” Journal of Experimental Medicine 198, no. 2 (2003): 305–313.

[42] M. Yarchoan, W. J. Ho, A. Mohan, et al., “Effects of B Cell-Activating Factor on Tumor Immunity,” JCI Insight 5, no. 10 (2020): e136417.

[43] H. Duan, L. Jing, X. Jiang, et al., “CD146 Bound to LCK Promotes T Cell Receptor Signaling and Antitumor Immune Responses in Mice,” Journal of Clinical Investigation 131, no. 21 (2021): e148568.

[44] J. Ampudia, Y.-G. WW, J. Badrani, et al., “CD6-ALCAM Signaling Regulates Multiple Effector/Memory T Cell Functions,” Journal of Immunology 204, no. supplement S1 (2020): 150.13.

[45] S. Tahara-Hanaoka, K. Shibuya, Y. Onoda, et al., “Functional Characterization of DNAM-1 (CD226) Interaction With Its Ligands PVR (CD155) and Nectin-2 (PRR-2/CD112),” International Immunology 16, no. 4 (2004): 533–538.

[46] L. Jiang, X. Fang, H. Wang, D. Li, and X. Wang, “Ovarian Cancer-Intrinsic Fatty Acid Synthase Prevents Anti-Tumor Immunity by Disrupting Tumor-Infiltrating Dendritic Cells,” Frontiers in Immunology 9 (2018): 2927.

[47] H. Yoon and S. Lee, “Fatty Acid Metabolism in Ovarian Cancer: Therapeutic Implications,” International Journal of Molecular Sciences 23, no. 4 (2022): 2170.

[48] B. Hu, J.-Z. Lin, X.-B. Yang, and X.-T. Sang, “Aberrant Lipid Metabolism in Hepatocellular Carcinoma Cells as Well as Immune Microenvironment: A Review,” Cell Proliferation 53, no. 3 (2020): e12772.

[49] B. Trabert, C. A. Hathaway, M. S. Rice, et al., “Ovarian Cancer Risk in Relation to Blood Cholesterol and Triglycerides,” Cancer Epidemiology and Prevention Biomarkers 30, no. 11 (2021): 2044–2051.

[50] K. A. Michels, T. S. McNeel, and B. Trabert, “Metabolic Syndrome and Risk of Ovarian and Fallopian Tube Cancer in the United States: An Analysis of Linked SEER-Medicare Data,” Gynecologic Oncology 155, no. 2 (2019): 294–300.

[51] A. J. Li, R. G. Elmore, C. IY-d, and B. Y. Karlan, “Serum Low-Density Lipoprotein Levels Correlate With Survival in Advanced Stage Epithelial Ovarian Cancers,” Gynecologic Oncology 116, no. 1 (2010): 78–81.

[52] Z. Zeng, F. Wei, and X. Ren, “Exhausted T Cells and Epigenetic Status,” Cancer Biology & Medicine 17, no. 4 (2020): 923–936.

[53] S. Li, S. Yang, and Y. Hong, “Higher Thymocyte Selection-Associated High Mobility Group Box (TOX) Expression Predicts Poor Prognosis in Patients With Ovarian Cancer,” BMC Cancer 22, no. 1 (2022): 1216.

[54] X. P. Lin, J. D. Mintern, and P. A. Gleeson, “Macropinocytosis in Different Cell Types: Similarities and Differences,” Membranes 10, no. 8 (2020): 177.

[55] V. Jayashankar and A. L. Edinger, “Macropinocytosis Confers Resistance to Therapies Targeting Cancer Anabolism,” Nature Communications 11, no. 1 (2020): 1121.

[56] C.-M. Högerkorp, S. Bilke, T. Breslin, S. Ingvarsson, and C. A. K. Borrebaeck, “CD44-Stimulated Human B Cells Express Transcripts Specifically Involved in Immunomodulation and Inflammation as Analyzed by DNA Microarrays,” Blood 101, no. 6 (2003): 2307–2313.

[57] Y. Liu, P. Chanana, J. I. Davila, et al., “Gene Expression Differences Between Matched Pairs of Ovarian Cancer Patient Tumors and Patient-Derived Xenografts,” Scientific Reports 9, no. 1 (2019): 6314.

[58] S. Chowdhury, J. J. Kennedy, R. G. Ivey, et al., “Proteogenomic Analysis of Chemo-Refractory High-Grade Serous Ovarian Cancer,” Cell 186, no. 16 (2023): 3476–3498.e35.

[59] V. K. Bhosle, T. Mukherjee, Y. W. Huang, et al., “SLIT2/ROBO1-Signaling Inhibits Macropinocytosis by Opposing Cortical Cytoskeletal Remodeling,” Nature Communications 11, no. 1 (2020): 4112.

[60] P. Karran, J. Offman, and M. Bignami, “Human Mismatch Repair, Drug-Induced DNA Damage, and Secondary Cancer,” Biochimie 85, no. 11 (2003): 1149–1160.

[61] D. L. Richardson, R. N. Eskander, and D. M. O'Malley, “Advances in Ovarian Cancer Care and Unmet Treatment Needs for Patients With Platinum Resistance: A Narrative Review,” JAMA Oncology 9, no. 6 (2023): 851–859.

[62] N. Ahmed and K. L. Stenvers, “Getting to Know Ovarian Cancer Ascites: Opportunities for Targeted Therapy-Based Translational Research,” Frontiers in Oncology 3 (2013): 256.

[63] K. Varga, Z.-J. Jiang, and L.-W. Gong, “Phosphatidylserine Is Critical for Vesicle Fission During Clathrin-Mediated Endocytosis,” Journal of Neurochemistry 152, no. 1 (2020): 48–60.

[64] L. E. Lupien, K. Bloch, J. Dehairs, et al., “Endocytosis of Very Low-Density Lipoproteins: An Unexpected Mechanism for Lipid Acquisition by Breast Cancer Cells,” Journal of Lipid Research 61, no. 2 (2020): 205–218.

[65] K. A. Sacksteder, J. C. Morrell, R. J. Wanders, R. Matalon, and S. J. Gould, “MCD Encodes Peroxisomal and Cytoplasmic Forms of Malonyl-CoA Decarboxylase and Is Mutated in Malonyl-CoA Decarboxylase Deficiency,” Journal of Biological Chemistry 274, no. 35 (1999): 24461–24468.

[66] M. De Martino, J. C. Rathmell, L. Galluzzi, and C. Vanpouille-Box, “Cancer Cell Metabolism and Antitumour Immunity,” Nature Reviews Immunology 24, no. 9 (2024): 654–669.

[67] S. Xu, O. Chaudhary, P. Rodríguez-Morales, et al., “Uptake of Oxidized Lipids by the Scavenger Receptor CD36 Promotes Lipid Peroxidation and Dysfunction in CD8+ T Cells in Tumors,” Immunity 54, no. 7 (2021): 1561–1577.e7.

[68] X. Ma, L. Xiao, L. Liu, et al., “CD36-Mediated Ferroptosis Dampens Intratumoral CD8+ T Cell Effector Function and Impairs Their Antitumor Ability,” Cell Metabolism 33, no. 5 (2021): 1001–1012.e5.

[69] K. Tondo-Steele and K. McLean, “The “Sweet Spot” of Targeting Tumor Metabolism in Ovarian Cancers,” Cancers 14, no. 19 (2022): 4696.

[70] W. Zhou, Y. Tu, P. J. Simpson, and F. P. Kuhajda, “Malonyl-CoA Decarboxylase Inhibition Is Selectively Cytotoxic to Human Breast Cancer Cells,” Oncogene 28, no. 33 (2009): 2979–2987.

[71] C. Commisso, S. M. Davidson, R. G. Soydaner-Azeloglu, et al., “Macropinocytosis of Protein Is an Amino Acid Supply Route in Ras-Transformed Cells,” Nature 497, no. 7451 (2013): 633–637.

[72] J. Ma, T. Chen, S. Wu, et al., “iProX: An Integrated Proteome Resource,” Nucleic Acids Research 47, no. D1 (2019): D1211–D1217.

[73] T. Chen, J. Ma, Y. Liu, et al., “iProX in 2021: Connecting Proteomics Data Sharing With Big Data,” Nucleic Acids Research 50, no. D1 (2022): D1522–D1527.

RIGHTS & PERMISSIONS

2026 The Author(s). MedComm published by Sichuan International Medical Exchange & Promotion Association (SCIMEA) and John Wiley & Sons Australia, Ltd.