Md Sufyan Ansari, Varnit Chauhan, Aashi Rai and Sujeet Kumar Singh, Areej Akhtar, Nisha Chaudhary, Reegina Tyagi, Divya, Kashif Husain, S. Sharma, Raquib Alam, Md Shakir, Mehak Pracha, Samreen Anjum, Mohd Nadeem, Prial Taneja, Md Imam Faizan, Iqbal Azmi, Aditya Ramdas Iyer, Pragya Gupta, Mehwish Nafiz, Shayan Ali, Insha Mohi Uddin, MA Javid, Zahoor Ahmad Bhat, Hamenth Kumar Palani, Amit Kumar Srivastava, Ulaganathan Mabalirajan, Vikram Mathews, Mohammad Husain, Sivaprakash Ramalingam, Gaurav Kharya, Tanveer Ahmad
The persistence of CAR T cells and antigen escape remain major barriers to durable therapeutic success in hematologic malignancies. Our study integrates AI-guided design with targeted protein degradation to overcome these challenges. Utilizing an in-silico library of CAR constructs followed by an in vitro screening, we developed a predictive model, CARMSeD, which forecasts constructs prone to self-activation and dysfunction. Optimized bispecific CD20/CD19 CAR T cells demonstrate superior persistence and anti-tumor efficacy. To further improve durability, the platform incorporates a PROTAC-based module that selectively degrades AKT3, promoting FOXO4-driven mitochondrial fitness, central memory differentiation, and reduced mTOR signaling. We extended this strategy to develop a trispecific CAR T platform co-expressing a secretable CD3/CD22 bispecific engager, achieving potent tumor eradication even in CD19/CD20-negative malignancies demonstrates efficacy across patient-derived leukemia samples and solid tumor models. Together, our study introduces a next-generation AI-guided CAR T strategy that integrates structure-based optimization and intracellular modulation to improve persistence, broaden antigen coverage, and ensure durable therapeutic efficacy.