Immunoinformatic approach for a novel protein in CAR-T cell therapy in hematological malignancies
Immunoinformatic approach for a novel protein in CAR-T cell therapy in hematological malignancies
Niloufar Moradi,1,*Hanieh Mohtashamiasl ,2Mohammad Salehi,3Mohammad reza Kalani,4
1. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran 2. Department of Medical Biotechnology, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran 3. Department of Medical Genetics, School of Advanced Technologies in Medicine, Golestan University of Medical Sciences, Gorgan, Iran 4. Principal Scientist of Molecular Cellular Biology and Protein Synthesis, Illinois, USA
Introduction: Cellular therapy has emerged as a key tool in the treatment of hematological malignancies. An advanced cell therapy known as chimeric antigen receptor T cell therapy (CAR T-cell therapy) has been approved by the United States Food and Drug Administration (FDA) as KYMRIAH by Novartis and YESCARTA by Gilead/Kite pharma in the year 2017. A chimeric receptor is composed of an extracellular antigen recognition site along with some co-stimulating and signaling domains. On the whole, it turns out to be one of the most potent receptors on T cells targeting a specific type of cancer cell based on its antigenic marker. CD19 CAR T-cell therapy is the first clinically approved therapy for lymphoma with remarkable results in complete remission of B cell lymphoblastic leukemia up to 90%. The high rate of effectiveness of the CAR T-cell therapy against B-ALL justifies the investigation and application of this therapy for fatal diseases like all types of hematological malignancies. The most critical aspect of chimeric receptor therapy is designing and building an artificial receptor that is specific to a given type of cancer.
Methods: The in silico technique is an appropriate model to investigate the integrity and effectiveness of the engineered chimeric receptor prior to commencing in vitro experiments followed by clinical trials. This computerized experimental study aids in predicting the molecular mechanism of chimeric protein and how it interacts with both ligands. We have anticipated various features of the chimeric protein in terms of qualitative analysis (structure, protein modelling, physiological properties) and functional analysis (antigenicity, allergenicity, its receptor-ligand binding ability, involving signaling pathways). Furthermore, the reliability and validation of the binding mode of the chimeric protein against receptors were performed through a complex molecular dynamics simulation for a 100 ns timeframe in an aqueous environment.
Results: The obtained simulation study showed that CD99 was a better approachable marker as compared to CD19 due to its better binding energy score and also binding conformations stability. The in silico results indicated that the designed antigen-binding site of the chimeric protein can recognize and bind to epitopes of CD19 and CD99 on the surfaces of both T and B cells. Furthermore, this chimeric protein possesses the same immunogenicity as the original protein as it was synthesized through the codon bias of a mammalian expression system. Thus, the proposed model of dual targeting anti-CD99−CD19-CAR protein represents a universal immunotherapy intervention for hematology cancer that is both novel and successful.
Conclusion: The main purpose of this study was to construct a unique receptor with two different antigen-binding domains on T cells that can mimic the role of TCR. It will be a more advanced approach to recognize and kill that particular tumor marker bearing leukemia and lymphoma cells. Many research organizations and scientists from all over the world have started investigating and developing various models of CAR T-cell therapy. Recently, we have reported about the different forms of CAR T- cells that are in the process of development. Amongst all CAR T-cell receptors targeting dual tumor markers (Bispecific CAR T-cell or Tandem CAR T-cell) can offer extremely effective antitumor therapy for patients with B cell & T cell malignancies.