Genetic Regulation in Human NK Cells: Insights into Immunological Diversity

Background

The immune system plays a central role in recognizing and eliminating cancer cells, and understanding the cellular and molecular mechanisms underlying this immune surveillance is fundamental to developing effective immunotherapies. Natural Killer (NK) cells are cytotoxic effector lymphocytes that play a crucial role in the innate immune response by eliminating tumor and virus-infected cells without prior sensitization. Their activity is tightly regulated by a balance between activating and inhibitory receptors expressed on their surface. Engagement of these receptors with ligands on target cells determines whether the NK cell becomes activated, leading to the release of cytotoxic granules and pro-inflammatory cytokines.

Previous research in our laboratory has demonstrated that a single nucleotide polymorphism (SNP) in the gene encoding the inhibitory receptor NKG2A influences NK cell responsiveness. This genetic variation was found to modulate NK cell-mediated immune responses, ultimately affecting patient outcomes during immunotherapy for acute myeloid leukemia (AML).

Preliminary Data

In single-cell analyses, integrating both mRNA and protein expression profiles of NK cells isolated from healthy donors carrying different NKG2A SNP genotypes, we have identified a candidate gene that may impact NK cell function. We will now investigate how the protein encoded by this gene impacts NK cell killing of tumor cells and were it is located in the cell.

Project Description

The aim of this Master’s project is to investigate how our protein of interest is influenced by a genetic variation in NKG2A, and how the transcription of this protein impacts NK cell function towards tumor cells.

Methods

Primary NK cells will be isolated from genotyped healthy donors and stratified based on NKG2A SNP status.

Gene and protein expression of the candidate gene will be assessed using qPCR, flow cytometry/Western blot, and possibly microscopy to determine genotype-dependent differences and subcellular localization.

To investigate functional relevance, gene knockdown experiments will be performed using siRNA. Knockdown efficiency will be validated at mRNA and protein levels.

NK cell function will be evaluated using:

·       CD107a degranulation assays

·       Flow-based cytotoxicity assays against tumor cell lines

·       Cytokine production analysis (e.g., IFN-γ)

Statistical analyses will assess genotype-dependent differences and the impact of gene perturbation on NK cell anti-tumor activity.

Expected Outcomes

This project will provide insight into how genetic variation in inhibitory receptor genes shapes NK cell biology. Understanding these mechanisms could contribute to the development of personalized immunotherapeutic strategies in cancer treatment.

Requirements

• Background in biotechnology or similar education
• Basic laboratory skills
• Interest in immune cell biology

Supervision

The project will be carried out in the Tumor Immunology (TIMM) Lab at Sahlgrenska Center for Cancer Research at Sahlgrenska Academy, University of Gothenburg, under the supervision of Elin Bernson.