OR7A10-engineered CAR-NK Cells Can Serve as A Highly Effective Therapy Against Solid Tumors

Chimeric Antigen Receptor (CAR)-Natural Killer (NK) cell therapy holds great promise for treating solid tumors. Still, its application remains limited due to poor infiltration, persistence, and resistance of CAR-NK cells within the tumor microenvironment (TME).

Recently, Sidi Chen and Lei Peng from Yale University published a research paper in Nature titled "OR7A10 GPCR engineering boosts CAR-NK therapy against solid tumours". To identify synergistic targets capable of enhancing CAR-NK cell efficacy, the study conducted an unbiased in vivo CRISPR activation (CRISPRa) screen, followed by a barcoded targeted in vivo open reading frame (ORF) screen in primary human CAR-NK cells. The study identified and comprehensively validated OR7A10, a G protein-coupled receptor (GPCR), as the optimal candidate.

Figure 1. In in vivo CRISPRa and barcoded ORF screening experiments, OR7A10 was identified as a key factor in enhancing the anti-tumor efficacy of CAR-NK cells. (YANG, Luojia, et al., 2026)

By engineering CAR-NK cells to encode OR7A10 cDNA—a method that bypasses CRISPR technology and utilizes a simple manufacturing strategy—the researchers enhanced their proliferation, activation, degranulation, cytokine production, death ligand expression, chemokine receptor expression, cytotoxicity, persistence, metabolic fitness, and resistance to the tumor microenvironment.

Furthermore, exhaustion was mitigated in primary human NK cells derived from multiple peripheral blood and umbilical cord blood donors. OR7A10-enhanced CAR-NK cells demonstrated robust in vivo efficacy across various solid tumor models. For instance, in an orthotopic breast cancer mouse model, a 100% complete response was achieved, along with long-term tumor control and survival benefits. These findings suggest that OR7A10-engineered CAR-NK cells can serve as a highly effective and mass-producible off-the-shelf therapy for solid tumors.

NK cells are cytotoxic lymphocytes with powerful capabilities for anti-tumor activity and the clearance of virus-infected cells. They can bypass Major Histocompatibility Complex (MHC) restrictions and prior immune stimulation. By recognizing gene-encoded ligands associated with oncogenic transformation, NK cells can target cancer cells with low mutational burdens or those lacking neoantigen presentation.

Adoptive CAR-NK cell therapy is relatively safe, carries almost no risk of Graft-Versus-Host Disease (GVHD) or Cytokine Release Syndrome (CRS), and is suitable for large-scale industrial production. These advantages have driven research into developing NK-cell-based therapies for solid tumors. As of 2025, over 1,200 clinical trials are evaluating NK cells, including more than 160 trials for CAR-NK cell therapies (ClinicalTrials.gov). In fact, trials have already demonstrated favorable results in treating hematological malignancies.

While CAR-NK cell therapy has immense potential for solid tumors, key challenges remain, including limited tumor infiltration, insufficient proliferation, and poor persistence within the tumor microenvironment (TME). Currently, various strategies are being investigated to overcome these limitations, including cytokine engineering and the knockout of inhibitory regulators such as CISH, NKG2A, HIF1A, CALHM2, or CREM33.

Although gene knockouts can enhance NK cell function, this approach relies on CRISPR-mediated gene editing, which increases the complexity of cell therapy manufacturing. An alternative strategy involves incorporating "enhancers"—genes with the function of driving overexpression—into the CAR construct, providing a simple and scalable solution for CAR-NK cell manufacturing.

This study performed in vivo CRISPRa screening on primary human CAR-NK cells, followed by a targeted small-scale screening of in vivo labeled ORFs, to identify genes that enhance anti-tumor activity in vivo upon overexpression (referred to as "super-enhancers" or "enhancers"). Through these experiments, the study identified a highly potent gene, OR7A10, which boosts CAR-NK cell function and demonstrates powerful anti-tumor effects in vivo.