Supplemental Figures and Legends from Follicle-Stimulating Hormone Receptor Is Expressed by Most Ovarian Cancer Subtypes and Is a Safe and Effective Immunotherapeutic Target
Year: 2023
Type: supplementary-materials
Abstract: Supplemental Figure 1. (A) Cytotoxicity of FSHCER or mock transduced T-cells of OVCAR-3 luciferase tumor cells measured by luciferase detection after co-culture of 18 hours. (B) Gating strategy followed for the flow cytometry based cytotoxicity assay. After gating on the singlet cells we could gate the tumor cells and exclude T-cells by FSC and SSC (confirmed by the no Tcell control) and measure the percentage of living cells as double negatives from Annexin-V and 7AAD. Supplemental Figure 2. (A) immunohistochemistry showing a section CAOV3 tumor stained with FSHR18 antibody followed by secondary anti-mouse (FSHR18) or secondary alone (Negative). 20x, scale bar: 1 um. (B) Western blot and qPCR showing FSHR expression of different ovarian cancer and breast cancer tumor cell lines. (C) Tumor volume of three ovarian patient-derived xenograft tumors grown in the flank of NOD-SCID mice (n=2 mice per tumor, one case-one control, single experiment) injected intratumorally with 10 million FSHCER or mock transduced T-cells (arrows mark time of T-cell injection). Common axis used for easier cross comparison. Supplemental Figure 3. (A) FSHR mRNA of FACS-sorted ID8-Defb29/Vegf-a/Fshr from the peritoneal cavity of orthotopic ID8-Defb29/Vegf-a/Fshr-bearing mice treated with either FSHCER, mock transduced T-cells or PBS. (B) Normalized real-time quantitative-PCR of FSHR expression in the human healthy ovarian tissue shown in Figure 3A, a human serous ovarian carcinoma specimen and 2 additional cell lines. (C) Absence of expression of the FSHR in untransduced ID8-Defb29/Vegf-a cells, compared to FSHR-transduced clones (40 PCR cycles).
Authors: Alfredo Perales-Puchalt, Nikolaos Svoronos, Melanie R. Rutkowski, Michael J. Allegrezza, Amelia J. Tesone, Kyle K. Payne, Jayamanna Wickramasinghe, Jenny M. Nguyen, Shane W. O'Brien, Kiranmai Gumireddy, Qihong Huang, Mark G. Cadungog, Denise C. Connolly, Julia Tchou, Tyler J. Curiel, Jose R. Conejo-Garcia
Language: English
Subfield: Reproductive Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4361967689