Data from SPP1⁺ Tumor-Associated Macrophages Drive Immunotherapy Resistance via CD8⁺ T-cell Dysfunction in Clear-Cell Renal Cell Carcinoma

Abstract Tumor-associated macrophages (TAM) are key regulators of tumor immunity. With advances in single-cell analyses, secreted phosphoprotein 1 (SPP1)–positive TAMs have been observed across multiple tumor sites. However, their clinical relevance and phenotypic characteristics in clear-cell renal cell carcinoma (ccRCC) have not been comprehensively delineated. Using patient-level data from two in-house cohorts (n = 355), we explored the relationship between SPP1⁺ TAM infiltration and therapeutic response and prognosis in ccRCC. Four publicly available datasets consisting of 1,741 patients with ccRCC were included for external validation. Cytometry by time-of-flight and flow cytometry were utilized to phenotype SPP1⁺ TAMs and establish their impact on CD8⁺ T cells. Furthermore, we established an ex vivo culture system to test the potential therapeutic value of targeting SPP1 alone and in conjunction with PD-1 inhibitors in ccRCC. We found that patients with high SPP1⁺ TAM infiltration exhibited worse response to immunotherapy and dismal prognosis in ccRCC. SPP1⁺ TAMs exhibited an immunosuppressive and protumor phenotype, and were related to impaired effector function and terminal differentiation of CD8⁺ T cells. Blockade of SPP1 mitigated the protumor tumor microenvironment and reinvigorated CD8⁺ T-cell function. Combining PD-1 blockade with SPP1 blockade boosted the expansion of CD8⁺ T cells and enhanced antitumor efficacy. Together, these data indicate that elevated infiltration of SPP1⁺ TAMs is related to worse response to immunotherapy and dysfunction of CD8⁺ T cells in ccRCC. We conclude that SPP1 may serve as a potential therapeutic target in ccRCC.