In Vitro Evaluation of CD276-CAR NK-92 Functionality, Migration and Invasion Potential in the Presence of Immune Inhibitory Factors of the Tumor Microenvironment.
Antigens, Neoplasm
/ immunology
B7 Antigens
/ genetics
CRISPR-Cas Systems
Cell Line, Tumor
Cell Movement
Cytotoxicity, Immunologic
Fibroblasts
/ metabolism
Gene Expression Regulation, Neoplastic
Humans
Hydrogen-Ion Concentration
Hypoxia
Immune System
Immunosuppression Therapy
Immunosuppressive Agents
Immunotherapy, Adoptive
/ methods
In Vitro Techniques
Killer Cells, Natural
/ cytology
Lactic Acid
/ metabolism
Lentivirus
/ genetics
Melanoma
/ immunology
Neoplasm Invasiveness
Receptors, Chimeric Antigen
/ genetics
Skin Neoplasms
/ immunology
Tumor Microenvironment
B7-H3
CD276
CRISPR/Cas9
NK-92
chimeric antigen receptor
immune therapy
melanoma
Journal
Cells
ISSN: 2073-4409
Titre abrégé: Cells
Pays: Switzerland
ID NLM: 101600052
Informations de publication
Date de publication:
26 04 2021
26 04 2021
Historique:
received:
04
03
2021
revised:
09
04
2021
accepted:
22
04
2021
entrez:
30
4
2021
pubmed:
1
5
2021
medline:
16
11
2021
Statut:
epublish
Résumé
Melanoma is the most lethal of all skin-related cancers with incidences continuously rising. Novel therapeutic approaches are urgently needed, especially for the treatment of metastasizing or therapy-resistant melanoma. CAR-modified immune cells have shown excellent results in treating hematological malignancies and might represent a new treatment strategy for refractory melanoma. However, solid tumors pose some obstacles for cellular immunotherapy, including the identification of tumor-specific target antigens, insufficient homing and infiltration of immune cells as well as immune cell dysfunction in the immunosuppressive tumor microenvironment (TME). In order to investigate whether CAR NK cell-based immunotherapy can overcome the obstacles posed by the TME in melanoma, we generated CAR NK-92 cells targeting CD276 (B7-H3) which is abundantly expressed in solid tumors, including melanoma, and tested their effectivity in vitro in the presence of low pH, hypoxia and other known factors of the TME influencing anti-tumor responses. Moreover, the CRISPR/Cas9-induced disruption of the inhibitory receptor NKG2A was assessed for its potential enhancement of NK-92-mediated anti-tumor activity. CD276-CAR NK-92 cells induced specific cytolysis of melanoma cell lines while being able to overcome a variety of the immunosuppressive effects normally exerted by the TME. NKG2A knock-out did not further improve CAR NK-92 cell-mediated cytotoxicity. The strong cytotoxic effect of a CD276-specific CAR in combination with an "off-the-shelf" NK-92 cell line not being impaired by some of the most prominent negative factors of the TME make CD276-CAR NK-92 cells a promising cellular product for the treatment of melanoma and beyond.
Sections du résumé
BACKGROUND
Melanoma is the most lethal of all skin-related cancers with incidences continuously rising. Novel therapeutic approaches are urgently needed, especially for the treatment of metastasizing or therapy-resistant melanoma. CAR-modified immune cells have shown excellent results in treating hematological malignancies and might represent a new treatment strategy for refractory melanoma. However, solid tumors pose some obstacles for cellular immunotherapy, including the identification of tumor-specific target antigens, insufficient homing and infiltration of immune cells as well as immune cell dysfunction in the immunosuppressive tumor microenvironment (TME).
METHODS
In order to investigate whether CAR NK cell-based immunotherapy can overcome the obstacles posed by the TME in melanoma, we generated CAR NK-92 cells targeting CD276 (B7-H3) which is abundantly expressed in solid tumors, including melanoma, and tested their effectivity in vitro in the presence of low pH, hypoxia and other known factors of the TME influencing anti-tumor responses. Moreover, the CRISPR/Cas9-induced disruption of the inhibitory receptor NKG2A was assessed for its potential enhancement of NK-92-mediated anti-tumor activity.
RESULTS
CD276-CAR NK-92 cells induced specific cytolysis of melanoma cell lines while being able to overcome a variety of the immunosuppressive effects normally exerted by the TME. NKG2A knock-out did not further improve CAR NK-92 cell-mediated cytotoxicity.
CONCLUSIONS
The strong cytotoxic effect of a CD276-specific CAR in combination with an "off-the-shelf" NK-92 cell line not being impaired by some of the most prominent negative factors of the TME make CD276-CAR NK-92 cells a promising cellular product for the treatment of melanoma and beyond.
Identifiants
pubmed: 33925968
pii: cells10051020
doi: 10.3390/cells10051020
pmc: PMC8145105
pii:
doi:
Substances chimiques
Antigens, Neoplasm
0
B7 Antigens
0
CD276 protein, human
0
Immunosuppressive Agents
0
Receptors, Chimeric Antigen
0
Lactic Acid
33X04XA5AT
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
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