Chimeric antigen receptor-based T-cell immunotherapy is a promising strategy for treatment of hematological malignant tumors;however,its efficacy towards solid cancer remains challenging.We therefore focused on develo...Chimeric antigen receptor-based T-cell immunotherapy is a promising strategy for treatment of hematological malignant tumors;however,its efficacy towards solid cancer remains challenging.We therefore focused on developing nanobody-based CAR-T cells that treat the solid tumor.CD105 expression is upregulated on neoangiogenic endothelial and cancer cells.CD105 has been developed as a drug target.Here we show the generation of a CD105-specific nanobody,an anti-human CD105 CAR-T cells,by inserting the sequences for anti-CD105 nanobody-linked standard cassette genes into AAVS1 site using CRISPR/Cas9 technology.Co-culture with CD105+target cells led to the activation of anti-CD105 CAR-T cells that displayed the typically activated cytotoxic T-cell characters,ability to proliferate,the production of pro-inflammatory cytokines,and the specific killing efficacy against CD105+target cells in vitro.The in vivo treatment with anti-CD105 CAR-T cells significantly inhibited the growth of implanted CD105+tumors,reduced tumor weight,and prolonged the survival time of tumor-bearing NOD/SCID mice.Nanobody-based CAR-T cells can therefore function as an antitumor agent in human tumor xenograft models.Our findings determined that the strategy of nanobody-based CAR-T cells engineered by CRISPR/Cas9 system has a certain potential to treat solid tumor through targeting CD105 antigen.展开更多
基金supported,in part,by grants from Project of National Natural Scientific Foundation of China(number 81773254)Programs for Changjiang Scholars and Innovative Research Team in University(number IRT_15R13)+6 种基金International Cooperation Project of the Misnistry of Science and Technology of China(number 2015 DP A31320)Project for Innovative Research Team in Guangxi Natural Science Foundation(2015G XNSFFA139001)Project for International Nanobody Research Center of Guangxi(number GuiKe-AD17195001)Partial support was provided by the NIH-NIEHS(RIVER Award)R35 ES030443-01the NIEHS Superfund Research Program P42 ES004699,Guangxi First-class Discipline Project for Pharmaceutical Sciences(number GXFCDP-PS-2018)National Key Research and Development Plan"Intergovernmental Cooperation in International Scientific and Technological Innovation"(number 2019YFE0117300)Guangxi Science and Technology Base and Talents Project(number GuiKe-AD20238062).
文摘Chimeric antigen receptor-based T-cell immunotherapy is a promising strategy for treatment of hematological malignant tumors;however,its efficacy towards solid cancer remains challenging.We therefore focused on developing nanobody-based CAR-T cells that treat the solid tumor.CD105 expression is upregulated on neoangiogenic endothelial and cancer cells.CD105 has been developed as a drug target.Here we show the generation of a CD105-specific nanobody,an anti-human CD105 CAR-T cells,by inserting the sequences for anti-CD105 nanobody-linked standard cassette genes into AAVS1 site using CRISPR/Cas9 technology.Co-culture with CD105+target cells led to the activation of anti-CD105 CAR-T cells that displayed the typically activated cytotoxic T-cell characters,ability to proliferate,the production of pro-inflammatory cytokines,and the specific killing efficacy against CD105+target cells in vitro.The in vivo treatment with anti-CD105 CAR-T cells significantly inhibited the growth of implanted CD105+tumors,reduced tumor weight,and prolonged the survival time of tumor-bearing NOD/SCID mice.Nanobody-based CAR-T cells can therefore function as an antitumor agent in human tumor xenograft models.Our findings determined that the strategy of nanobody-based CAR-T cells engineered by CRISPR/Cas9 system has a certain potential to treat solid tumor through targeting CD105 antigen.
