Redirected T Cell Therapy to Cure Invasive Fungal Infections

项目摘要 嵌合抗原受体（CAR）T细胞疗法给耐药感染性疾病患者带来新希望 微生物，如曲霉、念珠菌或毛霉。这是第一次模式识别受体 （Dectin-1）已被改造为重定向T细胞特异性以控制真菌感染。Dectin-1 CAR（D-CAR）可以 激活细胞溶解机制，并可能激活穿孔素/颗粒酶和颗粒溶素途径， T细胞从D-CAR+ T细胞产生IFN-γ可以进一步增强对侵袭性真菌的先天免疫。 如果重组IFN-γ直接或来源于CD 4+辅助T细胞或天然IFN-γ， 杀伤细胞 在R21阶段，限制CAR T细胞疗法立即临床应用的两个主要因素将是 解决了：（1）快速增殖的β-葡聚糖特异性D-CAR+ T细胞的产生和（2）长期体内 持续控制侵袭性真菌感染。 几种类型的汽车目前用于临床试验以控制B细胞恶性肿瘤。因为它尚未 显然，CAR设计为给定患者提供完全胜任的T细胞活化，我们已经开发了一种 用于筛选多个CAR分子的方法。我们的团队开发了EZ-CAR平台， 通过混合和匹配来源于已知T细胞活化受体的组分，同时保持 靶向结构域完好无损使用这种方法，我们将用Dectin-1真菌产生大约21个D-CAR。 靶向结构域。快速生产（从供体收集PBMC后10天内）可改善治疗效果。 由于它避免了复制介导的T细胞衰老和终末衰老， 这种分化与体内持久性的丧失有关。 在R33阶段，研究将扩展到针对各种临床重要的机会性霉菌 （毛霉属、赛多孢子菌属）和酵母（念珠菌属）。MD安德森临床试验中鉴定的耐药分离株 实验室将用于验证D-CAR+ T细胞的治疗功效。在一些真菌中，如 根霉（毛霉目），β-葡聚糖层被糖胺聚糖（GAG）层掩盖。D-CAR+ T- 细胞疗法将与真菌细胞壁生物合成抑制剂如卡泊芬净组合使用 - 糖胺聚糖层，其将允许更好地识别和激活D-CAR+ T细胞疗法。 总之，由于原发性免疫缺陷，如遗传性免疫缺陷， 突变和继发性免疫缺陷，如人类免疫缺陷病毒感染，癌症， 移植很可能受益于免疫辅助治疗。开发单一工程T- 可以靶向各种病原体的细胞，如D-CAR+ T细胞，其将T细胞特异性重定向至 曲霉属、念珠菌属和毛霉属，是高度保证打击侵袭性真菌感染， 免疫功能低下的患者。

Project Summary Chimeric antigen receptor (CAR) T-cell therapy give new hope to patients suffering from drug-resistant infectious organisms such as Aspergillus, Candida, or Mucor. This is the first time that a pattern-recognition receptor (Dectin-1) has been adapted to redirect T-cell specificity to control fungal infection. Dectin-1 CAR (D-CAR) can activate the cytolytic machinery, and likely the perforin/granzyme and granulysin pathway, of genetically modified T-cells. The production of IFN- from the D-CAR+ T-cells may further augment innate immunity to invasive fungal infections if recombinant IFN-γ is administered pharmacologically or derived from CD4+ helper T-cells or natural killer cells. In the R21 phase, 2 major factors that limit immediate clinical applications of CAR T-cell therapy will be addressed: (1) generation of rapidly proliferative β-glucan-specific D-CAR+ T-cells and (2) long-term in vivo persistence to control invasive fungal infection. Several types of CARs are currently used in clinical trials to control B-cell malignancy. Because it is not yet apparent which CAR design provides fully competent T-cell activation for a given patient, we have developed an approach for screening multiple CAR molecules. Our team has developed the EZ-CAR platform for generating multiple CARs by mixing and matching components derived from known T-cell activating receptors while keeping the targeting domain intact. Using this approach, we will generate about 21 D-CARs with the Dectin-1 fungal targeting domain. Rapid production (within 10 days of PBMC collection from donor) may improve the therapeutic potential of the manufactured T-cells because it avoids the replication-mediated T-cell senescence and terminal differentiation that is associated with loss of in vivo persistence. In the R33 phase, the study will be expanded to target a wide variety of clinically important opportunistic molds (Mucor, Scedosporium) and yeasts (Candida). Drug-resistant isolates identified in MD Anderson clinical laboratories will be used for validating the therapeutic efficacy of the D-CAR+ T cells. In some fungi, such as Rhizopus (Mucorales family), the β-glucan layer is masked by the glycosaminoglycans (GAG) layer. D-CAR+ T- cell therapy will be used in combination with fungal cell wall biosynthesis inhibitors such as caspofungin to disrupt the glycosaminoglycans layer, which will allow better recognition and activation of the D-CAR+ T-cell therapy. In summary, patients suffering from invasive fungal infections due to primary immunodeficiencies such as genetic mutations and secondary immunodeficiencies such as human immunodeficiency virus infection, cancer, and transplantation are highly likely to benefit from immune adjuvant therapy. Development of single-engineered T- cells that can target various pathogens, such as D-CAR+ T-cells cells, which redirect T-cell specificity to Aspergillus, Candida, and Mucor species, is highly warranted to combat invasive fungal infections in immunocompromised patients.