A differentiated and durable allogeneic strategy applicable to cell therapies.

Area of Science:

• Immunotherapy
• Gene Editing
• Regenerative Medicine

Background:

• Autologous T-cell therapies show promise but face manufacturing challenges.
• Individualized production limits the widespread use of T-cell therapies.

Purpose of the Study:

• To develop a persistent, "off-the-shelf" allogeneic (Allo) T-cell therapy.
• To overcome limitations of autologous T-cell manufacturing.

Main Methods:

• Utilized a multiplex Nme2Cas9 base editor to knockout human leukocyte antigen (HLA) class I and II alleles (HLA-A, HLA-B, CIITA) while retaining HLA-C.
• Employed CRISPR/Cas9 and adeno-associated virus for site-specific CAR or TCR integration into the TRAC locus.
• Optimized T-cell engineering with orthogonal CRISPR/Cas9 cleavage, base editors, and lipid nanoparticle delivery.

Main Results:

• Achieved efficient production of allogeneic CAR T-cells (Allo-CAR T) with high editing rates and scalability.
• Demonstrated comparable functional activity of allogeneic T-cells to autologous counterparts in preclinical assays.
• Minimized chromosomal aberrations using the gene-editing approach.

Conclusions:

• Matching residual HLA-C alleles prevented donor T-cell rejection by host T and NK cells.
• The approach prevents graft-versus-host disease by removing endogenous TCR.
• Allogeneic T-cell therapy and gene-edited iPSCs (TKO) show potential for regenerative medicine applications.