BaEV-MLV Chimeric Lentiviral Vectors for Quiescent HSC Transduction
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Solution Overview
Problem
Current lentiviral vectors face challenges in efficiently transducing quiescent hematopoietic stem cells and lymphocytes due to their non-permissive nature, leading to low gene transfer rates and risks of multi-copy integration and genotoxicity, especially with VSV-G pseudotyped vectors which are sensitive to human complement and induce immune responses.
Innovation Solution
Pseudotyped lentiviral vectors with a chimeric envelope glycoprotein comprising the transmembrane and extracellular domain of the baboon endogenous retrovirus (BaEV) envelope glycoprotein fused with the cytoplasmic tail domain of the murine leukemia virus (MLV) envelope glycoprotein, or a modified BaEV envelope glycoprotein lacking the fusion inhibitory R peptide, enable efficient and stable transduction of hematopoietic cells, including quiescent HSCs and lymphocytes, at low vector doses with mild cytokine stimulation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If conventional lentiviral vectors (VSV-G pseudotyped) are used for transduction, then transduction efficiency in activated cells is improved, but transduction efficiency in quiescent HSCs deteriorates
Solution Approach 1:
The patent changes the envelope glycoprotein parameter from VSV-G to BaEV (baboon endogenous retrovirus) envelope glycoprotein. This parameter change enables the lentiviral vectors to transduce quiescent HSCs efficiently without requiring cell activation, thereby resolving the contradiction between transduction efficiency in activated cells and transduction capability in quiescent cells.
2Productivity
If high doses of VSV-G-LVs are used to achieve efficient transduction, then transduction efficiency is improved, but sensitivity to human complement and immune response increases
Solution Approach 1:
The patent changes the envelope glycoprotein parameter from VSV-G to BaEV envelope glycoprotein. This parameter change confers resistance to human complement system while maintaining high transduction efficiency at lower vector doses, thereby resolving the contradiction between transduction efficiency and complement sensitivity.
3Productivity
If lentiviral vectors are used to transduce quiescent HSCs, then gene transfer is achieved, but risk of multi-copy integration and genotoxicity increases
Solution Approach 1:
The patent changes the envelope glycoprotein parameter from VSV-G to BaEV envelope glycoprotein and optimizes transduction conditions (low MOI, mild cytokine stimulation). This parameter change enables efficient single-copy integration in quiescent HSCs without requiring cell activation, thereby resolving the contradiction between gene transfer efficiency and safety regarding multi-copy integration.
4Ease of manufacture
If VSV-G pseudotyped vectors are used, then ease of manufacture is improved, but stability in human serum and resistance to complement deteriorates
Solution Approach 1:
The patent changes the envelope glycoprotein parameter from VSV-G to BaEV envelope glycoprotein. This parameter change confers stability in human serum and resistance to complement system while maintaining ease of manufacture through standard pseudotyping protocols, thereby resolving the contradiction between ease of manufacture and stability in human serum.
Data Source
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AI summary
The present invention concerns a pseudotyped viral vector particle for transferring biological material into cells, wherein said vector particle comprises at least: - a chimeric envelope glycoprotein which comprises or consists in a fusion of the transmembrane and extracellular domain of a baboon endogenous retrovirus (BaEV) envelope glycoprotein and the cytoplasmic tail domain of a murine leukemia virus (MLV) envelope glycoprotein; or - a modified BaEV envelope glycoprotein wherein the cytoplasmic tail domain is devoid of the fusion inhibitory R peptide.