Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma
Background Cell therapies for solid tumors are thwarted by the hostile tumor microenvironment (TME) and by heterogeneous expression of tumor target antigens. We address both limitations with a novel class of chimeric antigen receptors based on plant lectins, which recognize the aberrant sugar residu...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMJ Publishing Group
2023-01-01
|
| Series: | Journal for ImmunoTherapy of Cancer |
| Online Access: | https://jitc.bmj.com/content/11/1/e005891.full |
| _version_ | 1797948486728024064 |
|---|---|
| author | Malcolm K Brenner Challice Bonifant Norihiro Watanabe Mary K McKenna Daniel Brenner Ada Ozcan Maureen Legendre Dafydd G Thomas Christopher Ashwood Richard D Cummings David M Markovitz |
| author_facet | Malcolm K Brenner Challice Bonifant Norihiro Watanabe Mary K McKenna Daniel Brenner Ada Ozcan Maureen Legendre Dafydd G Thomas Christopher Ashwood Richard D Cummings David M Markovitz |
| author_sort | Malcolm K Brenner |
| collection | DOAJ |
| description | Background Cell therapies for solid tumors are thwarted by the hostile tumor microenvironment (TME) and by heterogeneous expression of tumor target antigens. We address both limitations with a novel class of chimeric antigen receptors based on plant lectins, which recognize the aberrant sugar residues that are a ‘hallmark’ of both malignant and associated stromal cells. We have expressed in T cells a modified lectin from banana, H84T BanLec, attached to a chimeric antigen receptor (H84T-CAR) that recognizes high-mannose (asparagine residue with five to nine mannoses). Here, we tested the efficacy of our novel H84T CAR in models of pancreatic ductal adenocarcinoma (PDAC), intractable tumors with aberrant glycosylation and characterized by desmoplastic stroma largely contributed by pancreatic stellate cells (PSCs).Methods We transduced human T cells with a second-generation retroviral construct expressing the H84T BanLec chimeric receptor, measured T-cell expansion, characterized T-cell phenotype, and tested their efficacy against PDAC tumor cells lines by flow cytometry quantification. In three-dimensional (3D) spheroid models, we measured H84T CAR T-cell disruption of PSC architecture, and T-cell infiltration by live imaging. We tested the activity of H84T CAR T cells against tumor xenografts derived from three PDAC cell lines. Antitumor activity was quantified by caliper measurement and bioluminescence signal and used anti-human vimentin to measure residual PSCs.Results H84T BanLec CAR was successfully transduced and expressed by T cells which had robust expansion and retained central memory phenotype in both CD4 and CD8 compartments. H84T CAR T cells targeted and eliminated PDAC tumor cell lines. They also disrupted PSC architecture in 3D models in vitro and reduced total tumor and stroma cells in mixed co-cultures. H84T CAR T cells exhibited improved T-cell infiltration in multicellular spheroids and had potent antitumor effects in the xenograft models. We observed no adverse effects against normal tissues.Conclusions T cells expressing H84T CAR target malignant cells and their stroma in PDAC tumor models. The incorporation of glycan-targeting lectins within CARs thus extends their activity to include both malignant cells and their supporting stromal cells, disrupting the TME that otherwise diminishes the activity of cellular therapies against solid tumors. |
| first_indexed | 2024-04-10T21:44:05Z |
| format | Article |
| id | doaj.art-dd3ceb49b32e483f92d49b7f902abc2c |
| institution | Directory Open Access Journal |
| issn | 2051-1426 |
| language | English |
| last_indexed | 2024-04-10T21:44:05Z |
| publishDate | 2023-01-01 |
| publisher | BMJ Publishing Group |
| record_format | Article |
| series | Journal for ImmunoTherapy of Cancer |
| spelling | doaj.art-dd3ceb49b32e483f92d49b7f902abc2c2023-01-18T16:30:09ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262023-01-0111110.1136/jitc-2022-005891Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stromaMalcolm K Brenner0Challice Bonifant1Norihiro Watanabe2Mary K McKenna3Daniel Brenner4Ada Ozcan5Maureen Legendre6Dafydd G Thomas7Christopher Ashwood8Richard D Cummings9David M Markovitz10Baylor College of Medicine Center for Cell and Gene Therapy, Houston, Texas, USAAff1 grid.39382.330000 0001 2160 926XBaylor College of MedicineCenter for Cell and Gene Therapy Houston TX USACenter for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas, USACenter for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas, USADepartment of Bioengineering, Rice University, Houston, Texas, USACenter for Cell and Gene Therapy, Baylor College of Medicine, Houston, Texas, USADepartment of Internal Medicine, University of Michigan, Ann Arbor, Michigan, USADepartment of Pathology, University of Michigan, Ann Arbor, Michigan, USADepartment of Surgery, Harvard Medical School, Boston, Massachusetts, USADepartment of Surgery, Harvard Medical School, Boston, Massachusetts, USADepartment of Internal Medicine, University of Michigan, Ann Arbor, Michigan, USABackground Cell therapies for solid tumors are thwarted by the hostile tumor microenvironment (TME) and by heterogeneous expression of tumor target antigens. We address both limitations with a novel class of chimeric antigen receptors based on plant lectins, which recognize the aberrant sugar residues that are a ‘hallmark’ of both malignant and associated stromal cells. We have expressed in T cells a modified lectin from banana, H84T BanLec, attached to a chimeric antigen receptor (H84T-CAR) that recognizes high-mannose (asparagine residue with five to nine mannoses). Here, we tested the efficacy of our novel H84T CAR in models of pancreatic ductal adenocarcinoma (PDAC), intractable tumors with aberrant glycosylation and characterized by desmoplastic stroma largely contributed by pancreatic stellate cells (PSCs).Methods We transduced human T cells with a second-generation retroviral construct expressing the H84T BanLec chimeric receptor, measured T-cell expansion, characterized T-cell phenotype, and tested their efficacy against PDAC tumor cells lines by flow cytometry quantification. In three-dimensional (3D) spheroid models, we measured H84T CAR T-cell disruption of PSC architecture, and T-cell infiltration by live imaging. We tested the activity of H84T CAR T cells against tumor xenografts derived from three PDAC cell lines. Antitumor activity was quantified by caliper measurement and bioluminescence signal and used anti-human vimentin to measure residual PSCs.Results H84T BanLec CAR was successfully transduced and expressed by T cells which had robust expansion and retained central memory phenotype in both CD4 and CD8 compartments. H84T CAR T cells targeted and eliminated PDAC tumor cell lines. They also disrupted PSC architecture in 3D models in vitro and reduced total tumor and stroma cells in mixed co-cultures. H84T CAR T cells exhibited improved T-cell infiltration in multicellular spheroids and had potent antitumor effects in the xenograft models. We observed no adverse effects against normal tissues.Conclusions T cells expressing H84T CAR target malignant cells and their stroma in PDAC tumor models. The incorporation of glycan-targeting lectins within CARs thus extends their activity to include both malignant cells and their supporting stromal cells, disrupting the TME that otherwise diminishes the activity of cellular therapies against solid tumors.https://jitc.bmj.com/content/11/1/e005891.full |
| spellingShingle | Malcolm K Brenner Challice Bonifant Norihiro Watanabe Mary K McKenna Daniel Brenner Ada Ozcan Maureen Legendre Dafydd G Thomas Christopher Ashwood Richard D Cummings David M Markovitz Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma Journal for ImmunoTherapy of Cancer |
| title | Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma |
| title_full | Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma |
| title_fullStr | Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma |
| title_full_unstemmed | Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma |
| title_short | Novel banana lectin CAR-T cells to target pancreatic tumors and tumor-associated stroma |
| title_sort | novel banana lectin car t cells to target pancreatic tumors and tumor associated stroma |
| url | https://jitc.bmj.com/content/11/1/e005891.full |
| work_keys_str_mv | AT malcolmkbrenner novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT challicebonifant novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT norihirowatanabe novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT marykmckenna novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT danielbrenner novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT adaozcan novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT maureenlegendre novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT dafyddgthomas novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT christopherashwood novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT richarddcummings novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma AT davidmmarkovitz novelbananalectincartcellstotargetpancreatictumorsandtumorassociatedstroma |