Journal: Cancer Immunology Research

Article Title: Secreted Fas Decoys Enhance the Antitumor Activity of Engineered and Bystander T Cells in Fas Ligand–Expressing Solid Tumors

doi: 10.1158/2326-6066.cir-22-0115

Figure Lengend Snippet: Figure 1. CAR T cells are susceptible to suppression by tumor-expressed FasL. T cells from healthy donor PBMCs were transduced with SFG-based retrovirus engineered to express a second-generation CAR specific for human PSCA. A, Transduction efficiency was determined by flow cytometry on day 5 after transduction. Shown in the histogram are data representative of a single donor and in the bar graph are summary data (mean SEM, n ¼ 8). Statistical significance was calculated using an unpaired t test. B, Cytolytic activity of nontransduced control T cells (NT) and CAR PSCA T cells (CAR) was tested in a 51Cr-release assay using 5103 PSCA– 293T cells and PSCAþ CAPAN-1 and CFPAC-1 pancreatic cancer cells (mean SEM, n ¼ 3). Statistical difference was calculated using an unpaired t test. C and D, Expression of Fas (CD95) on T cells at baseline, upon activation, and after CAR transduction was determined by flow cytometry. Expression by T cells from a representative donor (C) and summary data (mean SEM, n ¼ 5; D). Significance was determined using one-way ANOVA. E, The viability of CAR PSCA T cells was determined by flow cytometry using Annexin V and 7-AAD staining after exposure to the indicated concentrations of recombinant FasL. The % viable cells (Annexin V–7-AAD–) under each condition is shown (mean SEM, n ¼ 3). Significance was determined using one-way ANOVA. F, FasL expression on CAPAN-1 and CFPAC-1 pancreatic cancer cell lines was assessed by flow cytometry during normal culture conditions and following 24-hour exposure to IFNg. Shown in the histograms are data representative of 3 independent analyses. G, CAPAN-1 tumor cells engineered to express PSCA-GFP were cultured with CAR PSCA T cells in the absence or presence of a Fas- blocking antibody(ZB4) before analysisby flow cytometry. Shown in the FACS plots are data representativeof a single sample and in the bar graph are summary data (mean SEM, n ¼ 4). Significance was determined using one-way ANOVA. H, Illustration demonstrating the susceptibility of CAR T cells to suppression by solid tumors via the Fas/FasL signaling axis. P > 0.05 ¼ nonsignificant (ns); , P ≤0.05; , P ≤0.01; , P ≤0.001, as indicated in the figure panels.

Article Snippet: Briefly, supernatant samples harvested from T cells activated in 24-well non-TC plates coated with recombinant PSCA antigen (CUSABIO; catalog no. CSB-YP018840HU) at indicated concentrations or irradiated K562-PSCA cells at indicated E:T. To measure the production of FDþ15 fusionmolecule, samples were used in soluble Fas and IL15 ELISAs (R&D Systems, catalog no. DY247) to measure the two components separately.

Techniques: Transduction, Cytometry, Activity Assay, Control, Release Assay, Expressing, Activation Assay, Staining, Recombinant, Cell Culture, Blocking Assay

Journal: Cancer Immunology Research

Article Title: Secreted Fas Decoys Enhance the Antitumor Activity of Engineered and Bystander T Cells in Fas Ligand–Expressing Solid Tumors

doi: 10.1158/2326-6066.cir-22-0115

Figure Lengend Snippet: Figure 2. Engineering a soluble Fas decoy receptor to sequester FasLCAR PSCA T cells were engineered to express Fas decoy (FD) by retroviral transduction and used for functional assessments. A, Diagram illustrating FD secreted by CAR T cells enhancing T-cell antitumor activity by blocking inhibitory FasL signaling. B, FD construct schematic is shown on top and CAR and FD coexpression in T cells from a representative donor assessed by flow cytometry on day 5 after transduction (density plots). C, Summary transduction data indicating expression of CAR PSCA and CAR.FD in T cells (mean SEM, n ¼ 8). D, Phenotype of CAR PSCA and CAR.FD T cells was compared 10 days after transduction by flow cytometry (mean SEM, n ¼ 3). No significant difference was observed between CAR PSCA and CAR.FD based on unpaired t test. E, Expansion of CAR PSCA and CAR.FD T cells stimulated with irradiated K562-PSCA tumor cells was monitored for six days using a flow cytometer (mean SEM, n ¼ 4). No significant difference was observed between CAR PSCA and CAR.FD based on an unpaired t test on days 3 and 6. F, Secretion of effector cytokines IFNg) and TNFa by CAR PSCA and CAR.FD T cells after 48-hour culture with K562 or K562-PSCA cells was measured using a multiplex cytokine assay (mean SEM, n ¼ 3). No significant difference was observed between CAR PSCA and CAR.FD based on an unpaired t test. G, Cytolytic function of CAR PSCA and CAR.FD T cells was compared in a chromium release assay using 293T, CAPAN-1, and CFPAC-1 cells as targets at indicated E:T (mean SEM, n ¼ 3). Unpaired t test indicated no significant difference between CAR PSCA and CAR.FD in tumor killing. H, FD secretion by CAR PSCA and CAR.FD T cells in the absence or presence of recombinant PSCA stimulation was measured in the supernatant by soluble Fas ELISA 48 hours after culture initiation (mean SEM, n ¼ 4). Statistical significance was calculated using an unpaired t test. P > 0.05 ¼ nonsignificant (ns); , P ≤0.05; , P ≤0.01; , P ≤0.001.

Article Snippet: Briefly, supernatant samples harvested from T cells activated in 24-well non-TC plates coated with recombinant PSCA antigen (CUSABIO; catalog no. CSB-YP018840HU) at indicated concentrations or irradiated K562-PSCA cells at indicated E:T. To measure the production of FDþ15 fusionmolecule, samples were used in soluble Fas and IL15 ELISAs (R&D Systems, catalog no. DY247) to measure the two components separately.

Techniques: Retroviral, Transduction, Functional Assay, Activity Assay, Blocking Assay, Construct, Cytometry, Expressing, Irradiation, Multiplex Assay, Cytokine Assay, Release Assay, Recombinant, Enzyme-linked Immunosorbent Assay

Journal: Cancer Immunology Research

Article Title: Secreted Fas Decoys Enhance the Antitumor Activity of Engineered and Bystander T Cells in Fas Ligand–Expressing Solid Tumors

doi: 10.1158/2326-6066.cir-22-0115

Figure Lengend Snippet: Figure 3. FD-engineered T cells exhibit superior function in the presence of FasL. FasL-neutralizing and T-cell function–enhancing properties of FD were assessed using in vitro immunoassays. A, FasL neutralization by FD was measured by incubating recombinant human FasL with fresh media, conditioned media obtained from 233T cells, or conditioned media obtained from 293T-FD cells and subsequently used in a human FasL ELISA (mean SEM, n ¼ 4). Statistical significance was calculated using one- way ANOVA. B and C, Suppression of FasL-mediated T-cell apoptosis by FD was assessed by measuring the viability of CAR PCSA T cells in different culture conditions—no FD or FasL [Control sup. (no FasL)]; no FD, in the presence of recombinant FasL [Control sup. (þ FasL)]; no FD in the presence of recombinant FasL and FasL-blocking antibody NOK-2 [Control sup. (þ FasL, þ NOK-2)]; recombinant FasL in the presence of FD [FD sup. (þ FasL)]. T-cell viability was measured by flow cytometry by Annexin V and 7-AAD staining after overnight culture. Flow cytometry plots demonstrating the viability of CAR PSCA T cells for a representative donor (B) and summary data (mean SEM, n ¼ 3) (C). Statistical significance was determined using one-way ANOVA. D and E, Antitumor activity of CAR PSCA and CAR.FD T cells was measured against CAPAN-1–PSCA tumor cells in a coculture assay (E:T of 1:10) and tumor as well as T-cell numbers were quantified by flow cytometry. Starting cell numbers (day 0) were used as reference to calculate fold change in cell numbers for all time points. Line graphs demonstrating fold change in tumor cell numbers (D) and fold change in T-cell numbers (mean SEM, n ¼ 4; E). Statistical significance was calculated using an unpaired t test. F, Pie chart depicting the percentage of polyfunctional (defined as cells producing 2 or more cytokines) CAR PSCA and CAR.FD T cells for a representative donor after 5-hour stimulation with CAPAN-1-PSCA cells (E:T of 1:4). Stimulated T cells were analyzed using IsoPlexis single-cell cytokine assay after stimulation. Numbers next to the pie charts indicate the number of cytokines produced by each fraction of T cells specified in the pie charts. Data representative of three independent donors. P > 0.05, nonsignificant (ns); , P ≤0.05; , P ≤0.01; , P ≤0.001.

Article Snippet: Briefly, supernatant samples harvested from T cells activated in 24-well non-TC plates coated with recombinant PSCA antigen (CUSABIO; catalog no. CSB-YP018840HU) at indicated concentrations or irradiated K562-PSCA cells at indicated E:T. To measure the production of FDþ15 fusionmolecule, samples were used in soluble Fas and IL15 ELISAs (R&D Systems, catalog no. DY247) to measure the two components separately.

Techniques: Cell Function Assay, In Vitro, Neutralization, Recombinant, Enzyme-linked Immunosorbent Assay, Control, Blocking Assay, Cytometry, Staining, Flow Cytometry, Activity Assay, Co-culture Assay, Cytokine Assay, Produced

Journal: Cancer Immunology Research

Article Title: Secreted Fas Decoys Enhance the Antitumor Activity of Engineered and Bystander T Cells in Fas Ligand–Expressing Solid Tumors

doi: 10.1158/2326-6066.cir-22-0115

Figure Lengend Snippet: Figure 4. FD and IL15 fusion protein (FDþ15) further enhance T-cell activity. T cells were retrovirally transduced to coexpress CAR PSCA and FDþ15 and then used in experiments to assess their function. A, Schematic diagram illustrating the FD and IL15 fusion (FDþ15) transgene. B and C, T-cell transduction to assess the expression of CAR PSCA, CAR PSCA, and FD, or CAR PSCA and FDþ15 was measured by flow cytometry 5 days after transduction. Flow cytometry density plots for a representative donor (B) and summary data indicating expression of the CAR PSCA, CAR.FD, and CAR.FDþ15 transgenes (mean SEM, n ¼ 8; C). D, Secretion of FDþ15 molecule by activated T cells was measured using Fas and IL15 ELISAs to separately measure the two components of the fusion molecule. Bar graphs demonstrate the concentration of Fas and IL15 in supernatant obtained from CAR.FDþ15 T cells cultured in the absence or presence of recombinant PSCA (mean SEM, n ¼ 4). Significance was calculated by unpaired t tests. E, Expansion of CAR PSCA, CAR.FD, and CAR.FDþ15 T cells stimulated with irradiated K562-PSCA tumor cells was monitored for 6 days using a flow cytometer (mean SEM, n ¼ 4). Statistical significance was calculated using one-way ANOVA. F and G, Antitumor activity of CAR PSCA, CAR.FD, and CAR.FDþ15 T cells was measured against CAPAN-1-PSCA tumor cells in a coculture assay (E:T of 1:10). Tumor as well as T-cell numbers were quantified by flow cytometry. Starting cell numbers (day 0) were used as a reference to calculate fold change in cell numbers for all time points. Graphs illustrate fold change in tumor cell numbers (F) and T-cell numbers (G) over time (mean SEM, n ¼ 4). Statistical significance was calculated using one-way ANOVA. P > 0.05 ¼ nonsignificant (ns); , P ≤0.05; , P ≤0.01; , P ≤0.001.

Article Snippet: Briefly, supernatant samples harvested from T cells activated in 24-well non-TC plates coated with recombinant PSCA antigen (CUSABIO; catalog no. CSB-YP018840HU) at indicated concentrations or irradiated K562-PSCA cells at indicated E:T. To measure the production of FDþ15 fusionmolecule, samples were used in soluble Fas and IL15 ELISAs (R&D Systems, catalog no. DY247) to measure the two components separately.

Techniques: Activity Assay, Transduction, Expressing, Cytometry, Flow Cytometry, Concentration Assay, Cell Culture, Recombinant, Irradiation, Co-culture Assay

Journal: Cancer Immunology Research

Article Title: Secreted Fas Decoys Enhance the Antitumor Activity of Engineered and Bystander T Cells in Fas Ligand–Expressing Solid Tumors

doi: 10.1158/2326-6066.cir-22-0115

Figure Lengend Snippet: Figure 5. Decoy-engineered CAR T cells demonstrate enhanced in vivo antitumor activity. In vivo function of decoy-engineered T cells was performed using the CAPAN-1 xenograft model in NSG mice. A, Schematic representation of the in vivo model for the assessment of the decoy-engineered T cells. 5 106 CAPAN-1-PSCA tumor cells were engrafted subcutaneously in NSG mice, which were subsequently (approximately 4 weeks after tumor implantation) treated with i.v. injection of T cells. T cells used in the study were labeled with firefly luciferase to facilitate in vivo imaging. B, Change in body weight was used as an indicator of symptoms of toxicity associated with T-cell treatment and was monitored in all mice used in the study (mean SEM, n ¼ 7 per group). No statistical differences were observed between different treatment groups according to two-way ANOVA. C, Representative mice images (3 mice shown) demonstrating localization and expansion of the nontransduced (NT), CAR PSCA, CAR.FD, CAR.FDþ15, and CAR.FDþ15 cells by bioluminescence imaging. D and E, Tumor volumes were measured using calipers once a week for all mice in the study and reported as measurements for individual animals (D) and summarized data for each treatment group (mean SEM, n ¼ 7; E). F, T-cell expansion and persistence at the tumor were monitored by weekly bioluminescence imaging. Summary of T-cell luminescence at the tumor is demonstrated for each treatment group (mean SEM, n ¼ 7). Statistical significance between different treatment groups was calculated using two- way ANOVA. P > 0.05 ¼ nonsignificant (ns); , P ≤0.05; , P ≤0.01; , P ≤0.001.

Article Snippet: Briefly, supernatant samples harvested from T cells activated in 24-well non-TC plates coated with recombinant PSCA antigen (CUSABIO; catalog no. CSB-YP018840HU) at indicated concentrations or irradiated K562-PSCA cells at indicated E:T. To measure the production of FDþ15 fusionmolecule, samples were used in soluble Fas and IL15 ELISAs (R&D Systems, catalog no. DY247) to measure the two components separately.

Techniques: In Vivo, Activity Assay, Tumor Implantation, Injection, Labeling, Luciferase, In Vivo Imaging, Imaging

Journal: Cancer Immunology Research

Article Title: Secreted Fas Decoys Enhance the Antitumor Activity of Engineered and Bystander T Cells in Fas Ligand–Expressing Solid Tumors

doi: 10.1158/2326-6066.cir-22-0115

Figure Lengend Snippet: Figure 6. Decoy-secreting cells promote the effector activity of bystander T cells. Bystander effects of decoy-engineered T cells were assessed in a CFPAC-1 xenograft tumor model in NSG mice using CAR MUC1 T cells as bystander cells. A, Illustration of the in vivo model for the assessment of bystander effects of decoy-engineered T cells. NSG mice were engrafted subcutaneously with CFPAC-1 tumor cells and approximately 4 weeks after tumor inoculation, were treated with an i.v. injection of T cells. To facilitate in vivo tracking of both T-cell components administered to mice, CAR MUC1 cells were labeled with CBG99, and CAR PSCA, CAR.FD, and CAR.FDþ15 cells were labeled with Akaluc luciferases, respectively. B, Bioluminescence images of two representative mice per treatment group demonstrating localization and expansion of indicated T cells by bioluminescence imaging. Bystander CAR MUC1 T cells are depicted on the left and CAR PSCA, CAR.FD, and CAR.FDþ15 cells on the right. C, Tumor volumes were measured using calipers once a week for all mice in the study and illustrated in the line graphs for individual animals. D, Summary of CAR MUC1 T-cell luminescence indicating expansion and persistence of these cells in mice treated with CAR MUC1þCAR PSCA, CAR MUC1þCAR.FD, or CAR MUC1þCAR.FDþ15 T cells (mean SEM, n ¼ 7). E, Summary tumor volume for all mice in each treatment group illustrated in 6C (mean SEM, n ¼ 7). Statistical significance in tumor volumes between different treatment groups was calculated using two-way ANOVA. P > 0.05 ¼ nonsignificant (ns); , P ≤0.05; , P ≤0.01; , P ≤0.001.

Article Snippet: Briefly, supernatant samples harvested from T cells activated in 24-well non-TC plates coated with recombinant PSCA antigen (CUSABIO; catalog no. CSB-YP018840HU) at indicated concentrations or irradiated K562-PSCA cells at indicated E:T. To measure the production of FDþ15 fusionmolecule, samples were used in soluble Fas and IL15 ELISAs (R&D Systems, catalog no. DY247) to measure the two components separately.

Techniques: Activity Assay, In Vivo, Injection, Labeling, Imaging

Journal: Oncoimmunology

Article Title: Co-stimulatory signaling determines tumor antigen sensitivity and persistence of CAR T cells targeting PSCA+ metastatic prostate cancer

doi: 10.1080/2162402X.2017.1380764

Figure Lengend Snippet: PSCA-BBζ CARs show antigen-dependent cytokine production in vitro . (a) Flow cytometric analysis of PSCA expression in human prostate cancer cell lines. DU145 and PC-3 cell lines were lentivirally transduced to over-express human PSCA under the control of the EF1α promoter (see materials and methods). PC-3-PGK100p cell line was generated by expressing human PSCA under the control of the indicated mutant PGK promoter (PGK100p). LAPC-9 cells endogenously express human PSCA. (b) Quantification of CD137 (top) and CD69 (bottom) expression on Mock, PSCA-28ζ, and PSCA-BBζ CAR T cells following a 1, 4, or 24 hour co-culture with the indicated tumor targets at a 1:2 effector:tumor (E:T) ratio. (c) IFNγ production quantified by ELISA in supernatants from PSCA-CAR T cells cultured overnight with DU145 or DU145-PSCA tumor cells. (d) Same as in (c) from PSCA-CAR T cells cultured overnight with PC-3, PGK100p, or PC-3-PSCA tumor cells. (e) IFNγ production quantified by ELISA in supernatants from PSCA-CAR T cells cultured overnight on plate-bound recombinant human PSCA at varying protein concentrations. (f) Representative FACS plots showing intracellular IFNγ and CD107a degranulation by PSCA-CAR T cells following a 4 – 6 hr co-culture with indicated tumor targets. (g) Quantification of intracellular IFNγ and CD107a degranulation by PSCA-CAR T cells from (f). Data are shown as n = 2 per group ± SD. All data are representative of at least two independent experiments.

Article Snippet: Varying concentrations of recombinant human PSCA protein (amino acids 23–95; Abnova) was coated overnight in 1X PBS at 4°C on high-affinity 96-well flat bottom plates (Corning).

Techniques: In Vitro, Expressing, Control, Generated, Mutagenesis, Co-Culture Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Recombinant

Journal: Biomedicines

Article Title: Differential Protein-Coding Gene Expression Profile in Patients with Prostate Cancer

doi: 10.3390/biomedicines12112509

Figure Lengend Snippet: Genes classified as drug targets in the DrugBank database. Protein-coding genes as targets for drugs related to the treatment of prostate cancer in all states (investigation, experimental, approved). In the case of target genes for many drugs, only the first 10 are reported.

Article Snippet: DB05933 , MK-4721 , A fully human monoclonal antibody directed to Agensys’ proprietary target Prostate Stem Cell Antigen (PSCA), an antigen expressed at significant levels on tumor cells from the majority of patients with all stages of prostate, pancreatic, and bladder cancers [investigation]. , PSCA [O43653].

Techniques: Diagnostic Assay, Positron Emission Tomography, Membrane, Imaging, Biomarker Discovery, Expressing, Derivative Assay

Journal: Frontiers in Immunology

Article Title: A Unique Cellular and Molecular Microenvironment Is Present in Tertiary Lymphoid Organs of Patients with Spontaneous Prostate Cancer Regression

doi: 10.3389/fimmu.2017.00563

Figure Lengend Snippet: Increased tumor antigen load and active production of plasma cells are found at late stages of prostate cancer . The 5-μm thick paraffin sections were stained with antibodies against proliferating cell nuclear antigen (PCNA) (red), prostate stem cell antigen (PSCA, green), and plasma cell antigen (PC, white) to visualize PSCA tumor antigen and plasma cell generation at different stages of cancer progression/regression. Representative 200× pictures were taken with a Zeiss Axioplan microscope and recorded with a Hamamatsu camera. (A) Cells with eccentric nuclei (white) and cells positive for PSCA are detected in tertiary lymphoid organs (TLO) of prostatic intraepithelial neoplasia (PIN) samples. (B) Plasma cells are detected on the margins of TLO from patients at intermediate stages of prostate cancer. (C) Active germinal center that contains proliferating plasma blasts, non-proliferating plasma cells, and PSCA + cells was found in a sample from a patient at advanced stages of prostate cancer. (D) Proliferating plasmablasts are detected in a small TLO from a patient with evanescent prostate carcinoma. (E) Low proliferation and scarce PSCA signal in the epithelium of a PIN patient. (F) Highly proliferative but still organized epithelium in a tumor area of a patient at intermediate stage of prostate cancer. (G) Disorganized and proliferating nests of epithelial cells in a tumor area of patient with advanced prostate cancer. (H) PSCA signal was drastically reduced and epithelium had minimal evidence of proliferation in samples from patients with evanescent prostate cancer. Yellow arrows are pointing to proliferating plasmablasts that are indicative of local antigenic stimulation, while white arrows are depicting PSCA + cells. Scale bar represents 100 μm.

Article Snippet: The primary antibodies were as follows: goat anti-human CD105 (AF1097, R&D Systems), rabbit anti cyclooxygenase 2 (GTX15191, GeneTex), mouse anti-human CD68 (clone PG-M1, GeneTex), goat anti-CD3 epsilon (clone M-20, Santa Cruz Biotechnology), rabbit anti-T bet (H-210, Santa Cruz biotechnology), rat anti-human Foxp3 (PCH101, eBioscience), goat-anti proliferating cell nuclear antigen (PCNA) (clone C-20, Santa Cruz Biotechnology), mouse anti-human Ki-67 (clone MIB-1, Dakocytomation), rabbit anti-human CD8 (clone SP16, Thermo Fisher Scientific), mouse anti-human CD20 (clone L-26, Abcam), rabbit anti-human PD-L1 (Invitrogen, PA5-28115), rabbit anti-IL17 (H-132, Santa Cruz Biotechnology), mouse anti-human CD21 (clone 2G9, Thermo Fisher Scientific), rat anti-peripheral node addressin (clone MECA-79, BD Pharmigen), rabbit anti-human CD138 (RB-9422-P1, Thermo Fisher Scientific), mouse anti-podoplanin (clone D2-40, GTX31231, GeneTex), rabbit anti-granzyme B (clone EPR8260, Abcam), rat anti-human DC-LAMP (clone 1010E1.01, Novus Biologicals), rabbit anti-human prostate stem cell antigen (PSCA) (GTX15168, GeneTex), rabbit anti-human CXCL10 (GTX31176, GeneTex), biotin-mouse anti-smooth muscle actin (clone 1A4, Thermo Fisher Scientific), and mouse anti-human plasma cell (clone LIV3G11, Thermo Fisher Scientific).

Techniques: Staining, Microscopy

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Prostate stem cell antigen (PSCA) expression in primary GC tissues and cell lines. a. Immunohistochemical staining for PSCA in normal gastric tissue and eight primary GC samples; scale bar = 100 μm. b. Detection of PSCA expression in three human GC cell lines, BGC-823, KATO III, and MKN-28 cells, by flow cytometry

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Expressing, Immunohistochemical staining, Staining, Flow Cytometry

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Generation of anti-prostate stem cell antigen (PSCA) CAR-T cells. a . The discrete CAR units of anti-PSCA CAR-T cells and GFP-T cells. b . Representative flow cytometric analyses of transfected T cells detected by flow cytometry. c . CCR7, CD62L, CD45RA, and CD45RO expression was detected on T cells after their generation

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Transfection, Flow Cytometry, Expressing

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Anti-PSCA CAR-T cells exhibited dramatic antitumor efficacy ex vivo. a . The lytic capacity towards different target cells, including BGC-823, KATO III and MKN-28 cells, was analyzed at the indicated effector-to-target ratios in a 24 h lysis assay. b . The concentrations of IL-2, IFN-γ, GM-CSF, and TNFα released by anti-PSCA CAR-T cells and GFP-T cells after coculture with BGC-823 cells overnight at an E:T ratio of 1:1 are shown. Error bars denote the s.e.m., and the results were calculated by an unpaired t test. * indicates p < 0.05; ** indicates p < 0.01; and *** indicates p < 0.001. c . Canonical T cell markers were detected by flow cytometry at a recommended E:T ratio of 1:1 after coculturing anti-PSCA CAR-T cells and GFP- T cells with target cell lines. d . Statistical analysis of three independent FACS results. Error bars denote the s.e.m.

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Ex Vivo, Lysis, Flow Cytometry

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Anti-PSCA CAR-T cells efficiently reduced tumor progression in BGC-823 models. a . Schematic representation depicting the time course of the experiment. b . Tumor volume was calculated according to the following formula: length × width 2 /2. c . Tumor weight of BGC-823 subcutaneously injected mice. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. d . Percentage of T cells in the tumor of the BGC-823 models. e . Percentage of T cells in the PB of the BGC-823 models and representative FACS plots of the i.v group and the p.t group, respectively. f . Percentage of T cells in the spleen of the BGC-823 models and representative FACS plots of the i.v group and the p.t group, respectively. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Injection

Journal: Biomarker Research

Article Title: PSCA is a target of chimeric antigen receptor T cells in gastric cancer

doi: 10.1186/s40364-020-0183-x

Figure Lengend Snippet: Anti-PSCA CAR-T cells suppressed tumor progression in MKN-28 models. a . Schematic representation depicting the time course of the experiment. b . Tumor volume was calculated according to the following formula: length × width 2 /2. c . Tumor weight of MKN-28 subcutaneously injected mice. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001. d . Percentage of T cells in the tumor of the MKN-28 models. e . Percentage of T cells in the PB of the MKN-28 models and representative FACS plots of the i.v group and the p.t group, respectively. f . Percentage of T cells in the spleen of the MKN-28 models and representative FACS plots of the i.v group and the p.t group, respectively. Error bars denote the s.e.m., and the results were compared with one-way ANOVA. * p < 0.05; ** p < 0.01; *** p < 0.001

Article Snippet: The anti-PSCA scFv fragment was derived from the humanized IG8 anti-PSCA antibody [ ] and was synthesized by Genscript Co., Ltd. (NanJing, China) after codon optimization.

Techniques: Injection

Journal: medRxiv

Article Title: Insights into Renal Protein Handling Through GWAS of the Human Urine Proteome

doi: 10.64898/2025.12.08.25341835

Figure Lengend Snippet: a , Distribution of dilution-corrected PSCA levels in urine, color-coded for samples with levels detected above and below the assay’s limit of detection (top) and for the urine donors’ rs71514093 genotypes (bottom). b , Receiver operating characteristic (ROC) curve for the prediction of rs71514093 genotypes (i.e. the presence of at least one TG allele) based on urine levels of PSCA. c , Association of rs71514093 with urine PSCA levels and other molecular and clinical traits. Traits were selected based on positive genetic colocalization (PP H4 >0.8) with the PSCA association signal in urine, indicating a shared causal variant. Effect size and odds ratio are shown for the TG allele. pGWAS: GWAS of protein levels; PheWAS: phenome-wide association study; UTI: urinary tract infection.

Article Snippet: Together, our data suggest that urine PSCA levels are an accurate readout of the PSCA-mediated genetic predisposition to cancers and infections of the urinary and gastrointestinal tract, where the protein is highly expressed (GTEx Project and Human Protein Atlas , ).

Techniques: Variant Assay, Infection