Journal: Molecular Cancer

Article Title: In vivo gene transfer targeting in pancreatic adenocarcinoma with cell surface antigens

doi: 10.1186/1476-4598-11-81

Figure Lengend Snippet: Anti-MUC4 antibodies bound to modified lentiviruses drive efficient gene transfer in vivo in CAPAN2 cells. (A) CAPAN2 cells were grafted subcutaneously in the right flanks of immune-deficient recipient mice. When tumors reached about 100 mm 3 , lentiviruses carrying the luciferase reporter gene combined with anti-MUC4, anti-CLDN18 (PDAC targeting antibodies), anti-HLA (positive control for the targeted gene transfer), rabbit IgGs (ISO, negative control for the targeted gene transfer) or packaged into the broad tropism VSV-G-containing envelope (positive control) were directly injected in the tumors (n = 3 in each group). Two weeks after virus injections luciferase signal was measured in anesthetized live animals with a photon imager apparatus. (B) Quantification of luminescence in photons/steradiant/s (ph/sr/s) was divided by tumor mass in mg, since tumor mass could be very different in individuals after resection, and is expressed as a percentage of VSV-G positive control. Red circles depict the zones used for quantification (same size for all the mice). The black arrow points the area measured for background signal determination. Student’s t tests against the VSV-G condition were statistically different for ISO (p = 0.04) and CLDN18 (p = 0.05). (C) Western-blot analysis of luciferase expression in the previous tumor extracts. The negative control (CTNEG) corresponds to a tumor extract of untransduced CAPAN2 cells. The positive control (CTPOS) was obtained from a tumor derived from CAPAN2 cells transduced with a lentivirus bearing the bicistronic transgene LUCIFERASE-IRES-ZsGREEN. Extracts obtained from tumors transduced by oncospecific lentiviruses are identified according to the target cell surface antigen. ISO: rabbit IgGs.

Article Snippet: CAPAN2 and BXPC3 were kindly provided by Joel Tardive-Lacombe (INSERM U624, Marseille, France).

Techniques: Modification, In Vivo, Luciferase, Positive Control, Negative Control, Injection, Virus, Western Blot, Expressing, Derivative Assay, Transduction

Journal: Biomarker Research

Article Title: Identification of GRIN2D as a novel therapeutic target in pancreatic ductal adenocarcinoma

doi: 10.1186/s40364-023-00514-4

Figure Lengend Snippet: GRIN2D was identified as an oncogene in PDAC. ( A ). Workflow of in-vivo genome-wide RNAi screening. ( B ). GRIN2D was upregulated in PDAC cells, as compared to non-tumor HPDE cells. ( C ). IHC staining of GRIN2D in blood vessels and ducts with the respective markers PECAM-1 and CK19. GRIN2D was upregulated in PDAC tumor tissues, compared to adjacent non- tumor tissues. ( D ). GRIN2D expression was increased in PDAC tumors, as compared to normal, from two independent samples cohort GSE16515 and GSE15471. Data are from at least three independent experiments. Mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: PDAC cell line Capan2 cells were transduced with lentivirus carrying a human genome-wide shRNA library (GeneNetTM Human 50 K siRNA Library, System Biosciences).

Techniques: In Vivo, Genome Wide, Immunohistochemistry, Expressing

Journal: Biomarker Research

Article Title: Identification of GRIN2D as a novel therapeutic target in pancreatic ductal adenocarcinoma

doi: 10.1186/s40364-023-00514-4

Figure Lengend Snippet: GRIN2D drove tumorigenic functions in PDAC cells. ( A ). GRIN2D was knocked down by siRNAs in SW1990 and PANC04.03 cells. ( B ). Knockdown of GRIN2D inhibited cell growth in PDAC cells. ( C ). Knockdown of GRIN2D inhibited colony formation in PDAC cells. ( D ). Knockdown of GRIN2D inhibited cell migration in PDAC cells. ( E ). Knockdown of GRIN2D inhibited cell invasion in PDAC cells. Cells in the colony formation assay and invasion assay were stained by crystal violet. Data are from at least three independent experiments. Mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: PDAC cell line Capan2 cells were transduced with lentivirus carrying a human genome-wide shRNA library (GeneNetTM Human 50 K siRNA Library, System Biosciences).

Techniques: Knockdown, Migration, Colony Assay, Invasion Assay, Staining

Journal: Biomarker Research

Article Title: Identification of GRIN2D as a novel therapeutic target in pancreatic ductal adenocarcinoma

doi: 10.1186/s40364-023-00514-4

Figure Lengend Snippet: GRIN2D promotes PDAC progression by CREB/ p38 MAPK signaling pathway. ( A ). p38, MSK, and CREB were dephosphorylated after knockdown of GRIN2D in SW1990 and PANC04.03 cells. ( B ). Clinical correlation between GRIN2D and p-CREB levels in PDAC primary tumors. ( C ). Amount of common differential genes in SW1990 and PANC04.03 cells after knockdown of GRIN2D. ( D ). Expression of differential genes after knockdown of GRIN2D. ( E ). Correlation between GRIN2D and IL20RB, CCL22, G5S2, IL6, EPGN, and HMGA2 expression in PDAC tumors. ( F ). IL20RB, CCL22, G5S2, IL6, EPGN, and HMGA2 expression in tumor and non- tumor groups from two independent samples cohort GSE16515 and GSE15471. ( G ). Survival analysis of IL20RB, CCL22, G5S2, IL6, EPGN, and HMGA2 in PDAC patients. ( H ). Phosphorylated CREB binding to HMGA2 and IL20RB in SW1990 and PANC04.03 cells, as revealed by ChIP assay. ( I ). EMT markers ZEB1, SNAIL, SLUG, and TWIST were downregulated after knockdown of GRIN2D in SW1990 and PANC04.03 cells. Data are from at least three independent experiments. Mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: PDAC cell line Capan2 cells were transduced with lentivirus carrying a human genome-wide shRNA library (GeneNetTM Human 50 K siRNA Library, System Biosciences).

Techniques: Knockdown, Expressing, Binding Assay

Journal: Biomarker Research

Article Title: Identification of GRIN2D as a novel therapeutic target in pancreatic ductal adenocarcinoma

doi: 10.1186/s40364-023-00514-4

Figure Lengend Snippet: GRIN2D promoted tumor growth and liver metastasis in PDAC. ( A ). GRIN2D was knocked down by stable lentiviral shRNA system in SW1990 cells. ( B ). Knockdown of GRIN2D inhibited tumor growth in SW1990 cells. Photographs of mice xenograft at day 28 after knockdown of GRIN2D in SW1990 cells. ( C ). Tumor volume and tumor mass of xenograft mice after knockdown of GRIN2D. ( D ). IHC staining of Ki67, H&E Staining in mice subcutaneous tumors after knockdown of GRIN2D. ( E ). Knockdown of GRIN2D inhibited liver metastasis in SW1990 cells

Article Snippet: PDAC cell line Capan2 cells were transduced with lentivirus carrying a human genome-wide shRNA library (GeneNetTM Human 50 K siRNA Library, System Biosciences).

Techniques: shRNA, Knockdown, Immunohistochemistry, Staining

Journal: Biomarker Research

Article Title: Identification of GRIN2D as a novel therapeutic target in pancreatic ductal adenocarcinoma

doi: 10.1186/s40364-023-00514-4

Figure Lengend Snippet: Memantine is a potential drug for PDAC therapy. ( A ). Dose-response curve of memantine in PDAC cells. ( B ). Cell growth was inhibited in memantine treated PDAC cells, as revealed by CCk8 cell viability assay. ( C ). Cell migration was inhibited in memantine treated PDAC cells, as revealed by wound healing cell migration assay. ( D ). Clonogenic ability was inhibited in memantine treated PDAC cells, the cells in the colony formation assay were stained by crystal violet. ( E ). Level of GRIN2D decreased in memantine treated PDAC cells. ( F ). p38, MSK, and CREB in p38 MAPK signaling pathway were dephosphorylated after treatment of memantine. Data are from at least three independent experiments. Mean ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: PDAC cell line Capan2 cells were transduced with lentivirus carrying a human genome-wide shRNA library (GeneNetTM Human 50 K siRNA Library, System Biosciences).

Techniques: Viability Assay, Migration, Cell Migration Assay, Colony Assay, Staining