Related Experiment Video
Updated: Jul 8, 2025
Advances in Human Induced Pluripotent Stem Cell-Derived Chimeric Antigen Receptor-Expressing Natural Killer Cells
Published on: February 14, 2025
Harnessing the Power of CAR-NK Cells: A Promising Off-the-Shelf Therapeutic Strategy for CD38-Positive Malignancies
Maryam Asadi1,2,3, Razie Kiani3, Vahid Razban1
1Department of Molecular Medicine, School of Advanced Medical Sciences and Technologies, Shiraz University of Medical Sciences, Shiraz, Iran.
Background:
CD38 is highly expressed on multiple myeloma (MM) cells and has been successfully targeted by different target therapy methods. This molecule is a critical prognostic marker in both diffuse large B-cell lymphoma and chronic lymphocytic leukemia.
Objective:
We have designed and generated an anti-CD38 CAR-NK cell applying NK 92 cell line. The approach has potential application as an off-the-shelf strategy for treatment of CD38 positive malignancies.
Methods:
A second generation of anti-CD38 CAR-NK cell was designed and generated, and their efficacy against CD38-positive cell lines was assessed in vitro. The PE-Annexin V and 7-AAD methods were used to determine the percentage of apoptotic target cells. Flow cytometry was used to measure IFN-γ, Perforin, and Granzyme-B production following intracellular staining. Using in silico analyses, the binding capacity and interaction interface were evaluated.
Results:
Using Lentivirus, cells were transduced with anti-CD38 construct and were expanded. The expression of anti-CD38 CAR on the surface of NK 92 cells was approximately 25%. As we expected from in silico analysis, our designed CD38-chimeric antigen receptor was bound appropriately to the CD38 protein. NK 92 cells that transduced with the CD38 chimeric antigen receptor, generated significantly more IFN-γ, perforin, and granzyme than Mock cells, and successfully lysed Daudi and Jurkat malignant cells in a CD38-dependent manner.
Conclusion:
The in vitro findings indicated that the anti-CD38 CAR-NK cells have the potential to be used as an off-the-shelf therapeutic strategy against CD38-positive malignancies. It is recommended that the present engineered NK cells undergo additional preclinical investigations before they can be considered for subsequent clinical trial studies.
Insights
Engineered NK-92 cells targeting CD38 demonstrated potent anti-cancer activity in vitro. These anti-CD38 CAR-NK cells show promise as an off-the-shelf therapy for CD38-positive cancers.
Area of Science:
- Immunology
- Cell Biology
- Oncology
Background:
- CD38 is a key prognostic marker and therapeutic target in hematologic malignancies like multiple myeloma, diffuse large B-cell lymphoma, and chronic lymphocytic leukemia.
- High CD38 expression on cancer cells presents a viable target for novel therapeutic strategies.
Purpose of the Study:
- To design and generate a chimeric antigen receptor (CAR)-engineered Natural Killer (NK) cell therapy targeting CD38.
- To evaluate the in vitro efficacy and potential of this anti-CD38 CAR-NK cell approach as an off-the-shelf treatment for CD38-positive malignancies.
Main Methods:
- Generation of a second-generation anti-CD38 CAR-NK cell line using the NK-92 cell line and lentiviral transduction.
- In vitro assessment of CAR-NK cell cytotoxicity against CD38-positive cell lines using apoptosis assays (PE-Annexin V/7-AAD).
- Measurement of effector functions (IFN-γ, Perforin, Granzyme-B) via intracellular staining and flow cytometry; in silico analysis of CAR binding.
Main Results:
- Anti-CD38 CAR expression achieved approximately 25% on NK-92 cells, with successful binding to CD38 protein confirmed by in silico analysis.
- Engineered NK-92 cells produced significantly higher levels of IFN-γ, perforin, and granzyme compared to control Mock cells.
- The anti-CD38 CAR-NK cells effectively lysed CD38-positive malignant cell lines (Daudi and Jurkat) in a CD38-dependent manner.
Conclusions:
- In vitro studies demonstrate that anti-CD38 CAR-NK cells possess the potential for an off-the-shelf therapeutic strategy against CD38-positive hematologic cancers.
- Further preclinical investigations are warranted to assess the safety and efficacy of these engineered NK cells prior to clinical trials.
