A5038589956- CAR-T cell therapy research
- Monoclonal and Polyclonal Antibodies Research
- Viral Infectious Diseases and Gene Expression in Insects
- Immune Cell Function and Interaction
- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- Immunotherapy and Immune Responses
- Toxin Mechanisms and Immunotoxins
- Transgenic Plants and Applications
- Biosimilars and Bioanalytical Methods
University of Helsinki
2023
Chimeric antigen receptor (CAR) T-cell immunotherapies for solid tumors face critical challenges such as heterogeneous expression. We characterized stage-specific embryonic antigen-4 (SSEA-4) cell-surface glycolipid a target CAR therapy. SSEA-4 is mainly expressed during embryogenesis but also found in several cancer types making it an attractive tumor-associated antigen. Anti-SSEA-4 CAR-T cells were generated and assessed preclinically vitro vivo antitumor response safety. effectively...
<p>Supplementary Figure S4. Anti-tumor response and survival in the long-term vivo study. A. Treatment groups used T cell doses. B. Representative BLI images of mice treated with PBS, 1M, 2M NT or CAR-T cells doses at indicated time points (days; d). (M=1x106). *Asterisks indicate sacrificed due to toxicities.</p>
<p>Supplementary Figure S3. IFN measurement and organ BLI of the short-term in vivo study. A. quantification by ELISA post-mortem mouse blood plasma. B. Representative images isolated organs from mice treated with PBS, NT-T, CAR-T cells at indicated T cell doses after 32 days. C. Organ quantification. N=3 for control. N=5 all groups, except N=2 NT-T 1M, N=4 2M. Mean ± SD. One-way ANOVA Tukey’s multiple comparison tests. (A) ***p=0.0005, ****p=0.0001. (C) *p=0.04, **p≤0.001 to PBS...
<p>Supplementary Figure legends</p>
<p>Supplementary Data S1. Pathology Report of the long-term survival study. Post-mortem lung, intestine and kidney tissue samples collected analyzed by Histowiz.</p>
<p>Supplementary Figure S1. SSEA-4 staining specificity and LDH cytotoxicity assay. A. immunohistochemistry in paraffin-embedded gastric cancer tumor microarrays. B. Upper panel sections were treated with standard xylene/ethanol series used paraffin embedded section treatment, whereas lower samples untreated. C. Consecutive of fresh frozen pancreatic patient stained anti-SSEA-4 antibody. The was blocked hexaose (middle panel) or SSEA-3 (right panel). D. Representative...
<p>Supplementary Figure S2. Short-term in vivo study, dose-response, and weight monitoring. A. Treatment groups used T cell doses. B. Weight monitoring the indicated mice groups. CAR-T 3M treated showed a trend of mild loss. (M=1x106).</p>
<p>Supplementary Figure S3. IFN measurement and organ BLI of the short-term in vivo study. A. quantification by ELISA post-mortem mouse blood plasma. B. Representative images isolated organs from mice treated with PBS, NT-T, CAR-T cells at indicated T cell doses after 32 days. C. Organ quantification. N=3 for control. N=5 all groups, except N=2 NT-T 1M, N=4 2M. Mean ± SD. One-way ANOVA Tukey’s multiple comparison tests. (A) ***p=0.0005, ****p=0.0001. (C) *p=0.04, **p≤0.001 to PBS...
<p>Supplementary Figure S1. SSEA-4 staining specificity and LDH cytotoxicity assay. A. immunohistochemistry in paraffin-embedded gastric cancer tumor microarrays. B. Upper panel sections were treated with standard xylene/ethanol series used paraffin embedded section treatment, whereas lower samples untreated. C. Consecutive of fresh frozen pancreatic patient stained anti-SSEA-4 antibody. The was blocked hexaose (middle panel) or SSEA-3 (right panel). D. Representative...
<p>Supplementary Data S1. Pathology Report of the long-term survival study. Post-mortem lung, intestine and kidney tissue samples collected analyzed by Histowiz.</p>
<p>Supplementary Figure S6. Expression of SSEA-4 in normal mouse tissues. Immunofluorescence detection (green) the indicated tissues healthy 10 weeks old (C57BL/6) using anti-SSEA-4 antibody.</p>
<p>Supplementary Figure S4. Anti-tumor response and survival in the long-term vivo study. A. Treatment groups used T cell doses. B. Representative BLI images of mice treated with PBS, 1M, 2M NT or CAR-T cells doses at indicated time points (days; d). (M=1x106). *Asterisks indicate sacrificed due to toxicities.</p>
<p>Supplementary Figure S2. Short-term in vivo study, dose-response, and weight monitoring. A. Treatment groups used T cell doses. B. Weight monitoring the indicated mice groups. CAR-T 3M treated showed a trend of mild loss. (M=1x106).</p>
<p>Supplementary Figure S5. Weight monitoring during the long-term in vivo study. Mice treated with 2M CAR-T dose showed pronounced weight loss and health deterioration at earlier time points than mice 1M dose. PBS, tumor-free CAR/NT-T 1M, NT-T groups did not show remained healthy duration of experiment. Unless otherwise indicated, are tumor-bearing. (M=1x106).</p>
<p>Supplementary Figure legends</p>
<p>Supplementary Figure S7. T cell detection in mouse tissues after the long-term survival study. Indicated were stained with HOECHST 33342 for nuclei (blue), anti-EGFR to detect CAR-T cells (middle panel; green) and anti-CD3 (right panel, white). Left panel shows a merged image of all three channels. Arrows highlight CD3 positive cells, while arrowheads erythrocytes an intense background staining.</p>
<p>Supplementary Figure S6. Expression of SSEA-4 in normal mouse tissues. Immunofluorescence detection (green) the indicated tissues healthy 10 weeks old (C57BL/6) using anti-SSEA-4 antibody.</p>
<p>Supplementary Figure S7. T cell detection in mouse tissues after the long-term survival study. Indicated were stained with HOECHST 33342 for nuclei (blue), anti-EGFR to detect CAR-T cells (middle panel; green) and anti-CD3 (right panel, white). Left panel shows a merged image of all three channels. Arrows highlight CD3 positive cells, while arrowheads erythrocytes an intense background staining.</p>
<p>Supplementary Figure S5. Weight monitoring during the long-term in vivo study. Mice treated with 2M CAR-T dose showed pronounced weight loss and health deterioration at earlier time points than mice 1M dose. PBS, tumor-free CAR/NT-T 1M, NT-T groups did not show remained healthy duration of experiment. Unless otherwise indicated, are tumor-bearing. (M=1x106).</p>
<div>Abstract<p>Chimeric antigen receptor (CAR) T-cell immunotherapies for solid tumors face critical challenges such as heterogeneous expression. We characterized SSEA-4 cell-surface glycolipid a target CAR-T cell therapy. is mainly expressed during embryogenesis but also found in several cancer types making it an attractive tumor-associated antigen. Anti-SSEA-4 cells were generated and assessed pre-clinically vitro vivo anti-tumor response safety. effectively eliminated...
<div>Abstract<p>Chimeric antigen receptor (CAR) T-cell immunotherapies for solid tumors face critical challenges such as heterogeneous expression. We characterized stage-specific embryonic antigen-4 (SSEA-4) cell-surface glycolipid a target CAR therapy. SSEA-4 is mainly expressed during embryogenesis but also found in several cancer types making it an attractive tumor-associated antigen. Anti-SSEA-4 CAR-T cells were generated and assessed preclinically <i>in vitro</i>...
<p>Supplementary Figure S7. T cell detection in mouse tissues after the long-term survival study. Indicated were stained with HOECHST 33342 for nuclei (blue), anti-EGFR to detect CAR-T cells (middle panel; green) and anti-CD3 (right panel, white). Left panel shows a merged image of all three channels. Arrows highlight CD3 positive cells, while arrowheads erythrocytes an intense background staining.</p>
<p>Supplementary Figure S6. Expression of SSEA-4 in normal mouse tissues. Immunofluorescence detection (green) the indicated tissues healthy 10 weeks old (C57BL/6) using anti-SSEA-4 antibody.</p>