CLL1 CAR-T Cell Design for Selective AML Targeting
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Solution Overview
Problem
Current CAR-T cell therapy for Acute Myeloid Leukemia (AML) faces challenges due to the fundamental differences in antigenic properties of myeloid cells, leading to bone marrow failure and a lack of effective treatments, limiting its progress.
Innovation Solution
Development of CLL1-targeted CAR-T cells (CLL1-CAR-T cells) with a chimeric antigen receptor (CAR) comprising a single-domain antibody, hinge region, transmembrane region, and intracellular signaling region, specifically designed to target CLL1, a type II transmembrane glycoprotein expressed on AML cells, using a structure that includes a signal peptide, CLL1-VHH-1 antibody, CD8a hinge, CD8a transmembrane, co-stimulatory domain 4-1BB, and activation domain CD3ζ.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If traditional chemotherapy and hematopoietic stem cell transplantation are used to treat AML, then treatment can be provided, but efficacy is low and prognosis remains poor
Solution Approach 1:
The patent replaces traditional mechanical/chemical treatment methods (chemotherapy, stem cell transplantation) with a biological immunotherapy system (CAR-T cells). The CAR-T cells are genetically engineered to express chimeric antigen receptors that specifically recognize and bind to AML cells, enabling targeted immune-mediated destruction of tumor cells while sparing normal hematopoietic cells.
Solution Approach 2:
The patent modifies the T cells by introducing foreign genes (CAR genes) that encode chimeric antigen receptors. This genetic parameter change transforms ordinary T cells into specialized anti-AML effector cells with enhanced specificity and cytotoxicity against CLL1+ AML cells, fundamentally changing the treatment mechanism from non-specific chemotherapy to targeted immunotherapy.
2Reliability
If CAR-T cell therapy is applied to AML, then tumor targeting capability is improved, but bone marrow failure occurs due to killing of normal myeloid cells
Solution Approach 1:
The patent applies the principle of local quality by designing the CAR receptor with high specificity for CLL1 antigen expressed on AML cells. The single-domain antibody component is engineered to recognize unique epitopes on CLL1+ AML cells while displaying minimal cross-reactivity with normal myeloid cells, thereby concentrating the therapeutic effect locally on tumor cells and sparing normal bone marrow.
Solution Approach 2:
The patent uses the CAR-T cells as an intermediary between the immune system and AML cells. The chimeric antigen receptor acts as a mediator that translates recognition of CLL1 antigen into targeted cytotoxic responses, enabling selective elimination of AML cells while preserving normal hematopoietic cells that do not express or express low levels of CLL1.
3Adaptability or versatility
If conventional CAR structure is used, then general immunotherapy function is achieved, but specificity for AML targeting is insufficient due to antigenic property differences
Solution Approach 1:
The patent segments the CAR structure into distinct functional domains: a single-domain antibody variable region (VHH) for antigen recognition, a hinge region for flexibility, a transmembrane domain for membrane anchoring, and intracellular signaling domains (CD3ζ and co-stimulatory domains) for T cell activation. This segmentation allows optimization of each domain for AML-specific targeting while maintaining overall CAR functionality.
Solution Approach 2:
The patent creates a composite CAR structure by combining elements from different sources: the single-domain antibody (VHH) from camelid heavy chain antibodies for antigen binding, human CD3ζ signaling domain for T cell activation, and human co-stimulatory domains (4-1BB or CD28) for sustained T cell responses. This composite design integrates multiple functions into a single receptor optimized for AML targeting.
Data Source
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AI summary
This disclosure discloses a CLL1-CAR-T cell, as well as their preparation methods and applications. Specifically, it reveals CLL1-CAR-T cell that contain a chimeric antigen receptor, which comprises a single-domain antibody, a hinge region, a transmembrane region, and an intracellular signaling region. The amino acid sequence of the single-domain antibody corresponds to positions 22-150 of SEQ ID No.1. The CLL1-VHH-1 CAR-T cells of this disclosure can effectively secrete the T-cell-specific effector molecule IFN-γ, specifically and efficiently kill CLL1+ target cells, and exhibit favorable in vivo anti-tumor activity. They not only significantly inhibit the proliferation of tumor cells in mice but also markedly prolong the survival time of mice. The CLL1-VHH-1 CAR-T cells of this disclosure demonstrate excellent anti-tumor capabilities and can be used for immunotherapy of diseases related to the CLL1 target, such as acute myeloid leukemia, presenting broad prospects for clinical applications.