Recombinant PD-L1 peptides and methods of use
Inventors
Adeyanju, Kaothara Oluwakemi • Medin, Jeffrey A. • Hill, Robert Blake • Lum, Lawrence G.
Assignees
Medical College of Wisconsin • UVA Licensing and Ventures Group • University of Virginia UVA
Publication Number
US-12006511-B2
Publication Date
2024-06-11
Expiration Date
2040-04-14
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Abstract
The present disclosure provides soluble truncated mutant programmed death-ligand 1 (PD-L1) peptides. Polynucleotides and vectors encoding the soluble truncated mutant PD-L1 peptides are also provided. Further, methods of using the soluble truncated mutant peptides to promote differentiation of CD4+ effector T (Th1) cells into Foxp3+ regulatory T (Treg) cells are provided.
Core Innovation
The invention relates to soluble truncated mutant programmed death-ligand 1 (PD-L1) peptides and their encoding polynucleotides and vectors. These mutant peptides are derived from a truncated version of human PD-L1 that omits the transmembrane domain, corresponding to amino acids 18-132. The mutant peptides possess specific amino acid substitutions, including but not limited to D26E, D122E, Y123W, R125M, Y56I, S117I, and V68I, which can be present individually or in combination to improve efficacy in directing CD4+ effector T (Th1) cells to differentiate into Foxp3+ regulatory T (Treg) cells.
The problem addressed by this invention centers on the increasing incidence of autoimmune diseases, like Type 1 diabetes (T1D), and the limitations of current therapeutic strategies that fail to restore immune tolerance. The breakdown of immune tolerance mechanisms in these diseases leads to a reduction of functional Treg cells and an increase in autoreactive effector T cells, resulting in the destruction of target tissues such as pancreatic β cells. Existing methods of generating Treg cells are inefficient and laborious, often taking weeks to expand limited populations of Treg cells ex-vivo.
The disclosure provides a solution by engineering, expressing, and using soluble mutant PD-L1 peptides that markedly enhance the conversion of Th1 cells to Treg cells compared to wild-type PD-L1. The mutant peptides can be presented as part of peptide-bead complexes, encoded by specific polynucleotides, or delivered via vectors to host cells. Methods are described for generating Treg cells in vitro by contacting Th1 cells with these mutant PD-L1 peptides or peptide-bead complexes, with implications for more effective Treg cell production and restoring immune tolerance.
Claims Coverage
The claims encompass seven principal inventive features centering on soluble mutant PD-L1 peptides, related nucleic acids, expression systems, and methods for generating regulatory T cells.
Soluble mutant PD-L1 peptide with defined mutations
A soluble mutant PD-L1 peptide consisting essentially of SEQ ID NO: 1, wherein the peptide comprises one or more mutations selected from the group consisting of D9E, D105E, Y106W, R108M, Y39I, S100I, V51I, and combinations thereof, with mutation positions numbered relative to SEQ ID NO: 1.
Polynucleotide encoding the soluble mutant PD-L1 peptide
A polynucleotide encoding the soluble mutant PD-L1 peptide, as described, encoding SEQ ID NO: 1 with one or more defined mutations.
Vector expressing the soluble mutant PD-L1 peptide
A vector capable of expressing the soluble mutant PD-L1 peptide, wherein the vector comprises a promoter operably connected to a polynucleotide encoding the soluble mutant PD-L1 peptide.
Host cell comprising the PD-L1 expression vector
A host cell comprising the vector capable of expressing the soluble mutant PD-L1 peptide.
Soluble mutant PD-L1 peptide-bead complex
A complex comprising the soluble mutant PD-L1 peptide and a bead, where the peptide is covalently or non-covalently attached to the bead.
Method for generating regulatory Foxp3+ T cells
A method of generating regulatory Foxp3+ T cells (Foxp3+ Tregs), comprising contacting T cells cultured in vitro with either the soluble mutant PD-L1 peptide or the soluble mutant PD-L1 peptide-bead complex in an amount effective to generate Treg cells, where the peptide comprises SEQ ID NO: 2, 3, or 4.
In summary, the inventive features claim recombinant soluble mutant PD-L1 peptides with specific mutations, the nucleic acids and vectors encoding them, host cells and peptide-bead complexes containing these peptides, and methods of using them to induce regulatory T cells.
Stated Advantages
The mutant PD-L1 peptides direct Th1 cells to differentiate into Treg cells with greater efficiency than wild-type PD-L1 fragments.
The invention provides improved methods for generating large quantities of Treg cells ex-vivo, addressing limitations of current techniques that are inefficient and time-consuming.
Documented Applications
In vitro generation of regulatory T cells (Foxp3+ Tregs) from CD4+ effector T (Th1) cells.
Prevention, reduction, or slowing the progression or development of autoimmune diseases, such as type 1 diabetes, by increasing Treg cells in subjects.
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