Ahn, Wonkyung Han, Jihoon Kim, Nayeon Hwang, Yeong Ha Kim, Wonjun Lee, Yeram Lee, Dong Yun Cheong, In Woo Han, Koohee Nam, Gi-Hoon Kim, In-San Lee, Eun Jung 2025-03-22T14:30:08Z 2025-03-22T14:30:08Z 2025-03-19 2025-07 0142-9612 https://pubs.kist.re.kr/handle/201004/152019 Among cancer therapies, immune checkpoint blockade (ICB) has emerged as a prominent approach, substantially enhancing anti-tumor immune responses. However, the efficacy of ICB is often limited in the absence of a preexisting immune response within the tumor microenvironment. Here, we introduce a novel hierarchical protein hydrogel platform designed to facilitate the formation of artificial tertiary lymphoid structures (aTLS), thereby improving ICB efficacy. Through the integration of self-assembling ferritin protein nanocages, rec1-resilin protein, and CP05 peptide, our hierarchical hydrogels provide a structurally supportive and functionally adaptive scaffold capable of on-demand self-repair in response to mild thermal treatments. The effective encapsulation of extracellular vesicles (EVs) via the CP05 peptide ensures the formation of aTLS with germinal center-like structures within the hierarchical hydrogel. We demonstrate that, combined with ICB therapy, EV-loaded hierarchical hydrogels also induce the TLS within the tumor, markedly promoting immune responses against ICBresistant tumor. This bioactive hydrogel platform offers a versatile tool for enhancing a broad range of immunotherapies, with potential applications extending beyond TLS to other frameworks that support complex tissue architectures. English Elsevier Science Inc. Hierarchical protein nano-crystalline hydrogel with extracellular vesicles for ectopic lymphoid structure formation Article 10.1016/j.biomaterials.2025.123166 1 Biomaterials, v.318 Biomaterials 318 Y scie scopus 001426198400001 2-s2.0-85217078147 Engineering, Biomedical Materials Science, Biomaterials Engineering Materials Science Article FOLLICULAR DENDRITIC CELLS B-CELLS IMMUNOTHERAPY INDUCTION FERRITIN BIOMATERIALS ANGIOGENESIS MECHANISM IMMUNITY SURVIVAL Protein nano-crystalline Hierarchical hydrogel Extracellular vesicles Artificial tertiary lymphoid structure Cancer immunotherapy