| RRC ID |
83332
|
| Author |
Nakazawa T, Morimoto T, Maeoka R, Yamada K, Matsuda R, Nakamura M, Nishimura F, Yamada S, Park YS, Tsujimura T, Nakagawa I.
|
| Title |
Characterization of HIF-1α Knockout Primary Human Natural Killer Cells Including Populations in Allogeneic Glioblastoma.
|
| Journal |
Int J Mol Sci
|
| Abstract |
Enhancing immune cell functions in tumors remains a major challenge in cancer immunotherapy. Natural killer cells (NK) are major innate effector cells with broad cytotoxicity against tumors. Accordingly, NK cells are ideal candidates for cancer immunotherapy, including glioblastoma (GBM). Hypoxia is a common feature of solid tumors, and tumor cells and normal cells adapt to the tumor microenvironment by upregulating the transcription factor hypoxia-inducible factor (HIF)-1α, which can be detrimental to anti-tumor effector immune cell function, including that of NK cells. We knocked out HIF-1α in human primary NK cells using clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein 9 (Cas9). Then, cellular characterizations were conducted in normoxic and hypoxic conditions. Electroporating two HIF-1α-targeting guide RNA-Cas9 protein complexes inhibited HIF-1α expression in expanded NK cells. HIF-1α knockout human NK cells, including populations in hypoxic conditions, enhanced the growth inhibition of allogeneic GBM cells and induced apoptosis in GBM-cell-derived spheroids. RNA-sequencing revealed that the cytotoxicity of HIF-1α knockout NK cells could be related to increased perforin and TNF expression. The results demonstrated that HIF-1α knockout human NK cells, including populations, enhanced cytotoxicity in an environment mimicking the hypoxic conditions of GBM. CRISPR-Cas9-mediated HIF-1α knockout NK cells, including populations, could be a promising immunotherapeutic alternative in patients with GBM.
|
| Volume |
25(11)
|
| Published |
2024-5-28
|
| DOI |
10.3390/ijms25115896
|
| PII |
ijms25115896
|
| PMID |
38892084
|
| PMC |
PMC11173110
|
| MeSH |
Apoptosis / genetics
Brain Neoplasms / genetics
Brain Neoplasms / immunology
Brain Neoplasms / metabolism
Brain Neoplasms / pathology
CRISPR-Cas Systems
Cell Line, Tumor
Cytotoxicity, Immunologic
Gene Knockout Techniques*
Glioblastoma* / genetics
Glioblastoma* / immunology
Glioblastoma* / metabolism
Glioblastoma* / pathology
Humans
Hypoxia-Inducible Factor 1, alpha Subunit* / genetics
Hypoxia-Inducible Factor 1, alpha Subunit* / metabolism
Killer Cells, Natural* / immunology
Killer Cells, Natural* / metabolism
Tumor Microenvironment / genetics
Tumor Microenvironment / immunology
|
| IF |
4.556
|
| Resource |
| Human and Animal Cells |
T98G(RCB1954)
U251(RCB0461) |
