抑制免疫细胞和癌细胞的糖酵解会减弱它们的活性，因此将免疫疗法与糖酵解抑制剂相结合是具有挑战性的。在此，提出了一种策略，通过使用PFK15（PFKFB3的抑制剂，糖酵解中的限速步骤）在癌细胞中抑制糖酵解，同时通过给予果糖-1,6-二磷酸（F16BP，在PFKFB3下游的一步骤）来恢复DCs的糖酵解和功能。为了证明该策略的可行性，使用磷酸钙化学方法制备了疫苗配方，其中包含F16BP、聚合物(IC)作为佐剂，以及磷酸化-TRP2肽抗原，并在免疫能力正常的雌性小鼠的具有挑战性的和已建立的YUMM1.1肿瘤中进行了测试。此外，为了测试该策略的多功能性，还开发了包含F16BP、聚合物(IC)作为佐剂和源自B16F10细胞的mRNA作为抗原的移植DC疗法，并在免疫能力正常的雌性小鼠的已建立的B16F10肿瘤中进行了测试。F16BP疫苗配方在体外和体内恢复了DCs，并显著改善了小鼠的生存率，并通过提高肿瘤内的Tc1和Tc17细胞产生细胞毒性T细胞（Tc）反应。总的来说，这些结果表明可以利用基于代谢物的配方来恢复DCs的糖酵解，即使存在糖酵解抑制剂，也可以用于产生免疫疗法。© 2023. Springer Nature Limited.

Inhibition of glycolysis in immune cells and cancer cells diminishes their activity, and thus combining immunotherapies with glycolytic inhibitors is challenging. Herein, a strategy is presented where glycolysis is inhibited in cancer cells using PFK15 (inhibitor of PFKFB3, rate-limiting step in glycolysis), while simultaneously glycolysis and function is rescued in DCs by delivery of fructose-1,6-biphosphate (F16BP, one-step downstream of PFKFB3). To demonstrate the feasibility of this strategy, vaccine formulations are generated using calcium-phosphate chemistry, that incorporate F16BP, poly(IC) as adjuvant, and phosphorylated-TRP2 peptide antigen and tested in challenging and established YUMM1.1 tumours in immunocompetent female mice. Furthermore, to test the versatility of this strategy, adoptive DC therapy is developed with formulations that incorporate F16BP, poly(IC) as adjuvant and mRNA derived from B16F10 cells as antigens in established B16F10 tumours in immunocompetent female mice. F16BP vaccine formulations rescue DCs in vitro and in vivo, significantly improve the survival of mice, and generate cytotoxic T cell (Tc) responses by elevating Tc1 and Tc17 cells within the tumour. Overall, these results demonstrate that rescuing glycolysis of DCs using metabolite-based formulations can be utilized to generate immunotherapy even in the presence of glycolytic inhibitor.© 2023. Springer Nature Limited.