Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Systemic administration of human mesenchymal stromal cells infected with polymer-coated oncolytic adenovirus induces efficient pancreatic tumor homing and infiltration

doi: 10.1016/j.jconrel.2019.04.040

Figure Lengend Snippet: Cancer cell killing effect and viral replication of oAd/RLX-PCDP complex-loaded hMSCs on various pancreatic cancer cells. (A) Oncolytic effect by PBS, naked oAd/RLX, oAd/RLX-loaded hMSCs, or oAd/RLX-PCDP-loaded hMSCs. Cells (AsPC-1, PANC-1, and MIA PaCa-2) were infected with naked oAd/RLX, oAd/RLX-loaded or oAd/RLX-PCDP-loaded hMSCs at various MOIs. After 4 (AsPC-1 and PANC-1) or 6 (MIA PaCa-2) days post-infection, cell viability was measured by MTT assay. For oAd/RLX groups, the cell viability solely accounts for the absorbance readout of respective cancer cell types. For oAd/RLX- and oAd/RLX-PCDP-loaded hMSC groups, the total cell viability (as determined by absorbance readout) accounts for both respective types of pancreatic cancer cells and surviving hMSC carrier. (B) Viral production. Cells (AsPC-1, PANC-1, and MIA PaCa-2) were infected with naked oAd/RLX, oAd/RLX-loaded hMSCs, or oAd/RLX-PCDP-loaded hMSCs. At 72 h post-infection, viral genomic copies were measured by real-time quantitative PCR. Results represent the mean ± SD of triplicate experiments. ***P < 0.001 versus oAd/RLX-loaded hMSC control.

Article Snippet: In vivo antitumor efficacy Pancreatic cancer xenograft model was established by injecting 5 × 10 6 AsPC-1 cells into the subcutaneous of 6 weeks-old female nude mice (n=8) (Charles River Inc., Wilmington, MA).

Techniques: Infection, MTT Assay, Real-time Polymerase Chain Reaction

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Systemic administration of human mesenchymal stromal cells infected with polymer-coated oncolytic adenovirus induces efficient pancreatic tumor homing and infiltration

doi: 10.1016/j.jconrel.2019.04.040

Figure Lengend Snippet: Tumor-tropic migratory properties of oAd/RLX-PCDP complex-loaded hMSCs. oAd/RLX, oAd/RLX-PEI, and oAd/RLX-PCDP complex were treated into hMSCs at 5 MOI. After 18 h, these cells were detached and co-cultured with pancreatic cancer cells (AsPC-1) into the transwell plate for 30 h. Then, the migrated cells were fixed and stained with hematoxylin and eosin solution. All conditions were done in quadruplicate and repeated in two separate experiments. ***P < 0.001 versus uninfected hMSC (negative control) or naked oAd/RLX.

Article Snippet: In vivo antitumor efficacy Pancreatic cancer xenograft model was established by injecting 5 × 10 6 AsPC-1 cells into the subcutaneous of 6 weeks-old female nude mice (n=8) (Charles River Inc., Wilmington, MA).

Techniques: Cell Culture, Staining, Negative Control

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Systemic administration of human mesenchymal stromal cells infected with polymer-coated oncolytic adenovirus induces efficient pancreatic tumor homing and infiltration

doi: 10.1016/j.jconrel.2019.04.040

Figure Lengend Snippet: In vivo antitumor efficacy of oAd/RLX-PCDP-loaded hMSCs in tumor-bearing mice. Pancreatic tumor xenograft model was established by injecting AsPC-1 cells (5 × 106) subcutaneously in nude mice (n=8). Following the confirmation of tumorigenesis, the treatments were systemically administered on 4 day intervals for total of 3 injections when the average tumor volumes reached 90 mm3. (A) Relative tumor volume of each treatment group at day 21 post treatment in respect to initial tumor volume from day 1. (B) Relative body weight of each treatment group in respect to PBS treatment group at 21 days post initial treatment. (C) Histological and immunohistochemical analysis of tumor tissues from mice treated with PBS, PCDP, hMSC, naked oAd/RLX, oAd/RLX-loaded hMSC, or oAd/RLX-PCDP-loaded hMSC. Tumor tissues were collected from mice at 3 days after the final treatment. Representative sections were stained with H & E or MT solution. TUNEL assay was performed to detect apoptosis, and the expression of E1A was assessed by immunohistochemistry. Data presented as mean ± SD. ***P < 0.001 versus PBS, PCDP, hMSC, naked oAd/RLX, and oAd/RLX-loaded hMSC.

Article Snippet: In vivo antitumor efficacy Pancreatic cancer xenograft model was established by injecting 5 × 10 6 AsPC-1 cells into the subcutaneous of 6 weeks-old female nude mice (n=8) (Charles River Inc., Wilmington, MA).

Techniques: In Vivo, Immunohistochemical staining, Staining, TUNEL Assay, Expressing, Immunohistochemistry

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: Antitumor activity of BiTP combinatorial chemotherapy in orthotopic PDAC models. (A) Treatment plan and schedules. (B) Schematic diagram of the KPC model. The approximate time frame of mouse tumor progression to borderline resectable (BR), locally advanced (LA), and terminal (TERM) are days 7, 14, and 21, respectively. (C) Representative tumor images of orthotopic PDAC mice receiving different therapies. (D) Tumor weights and volumes on day 30. (E) The table included the survival rate (%), tumor proliferation (T/C, %), tumor growth inhibition (TGI, %), and resectability status of orthotopic PDAC mice receiving different therapies on day 30, and the median survival time (MST) in survival assays. (F) Kaplan–Meier plot survival curve and pairwise comparison results of orthotopic PDAC mice receiving different therapies in survival assays. Data presented in the graphs represent mean±standard deviation (SD) (**** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05; ns P >0.05). BiTP, anti-PD-L1/TGF-β antibody; CTH, chemotherapy; PDAC, pancreatic ductal adenocarcinoma; PD-L1, programmed cell death 1 ligand 1.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: Activity Assay, Inhibition, Comparison, Standard Deviation

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: Decisions about resectability status in PDAC murine models.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: Membrane

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: Preclinical assessment of neoadjuvant BiTP combinatorial chemotherapy for the treatment of PDAC. (A) Treatment plan and schematic diagram of treatment schedules. (B) Schematic diagram of distal pancreatectomy in orthotopic PDAC mice model. (C) Representative images of tumors. (D) Tumor weights and volumes of direct surgery (day 7) and post-neoadjuvant therapy resections (day 24). (E) Table of resectability status (day 24), operative mortality (OM), and median survival time (MST) of orthotopic PDAC mice receiving different therapies. (F–H) Kaplan–Meier plot survival curve and pairwise comparison results of orthotopic PDAC mice receiving different therapies in survival assays. Data presented in the graphs represent mean±standard deviation (SD). (**** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05; ns P >0.05). AC, adjuvant chemotherapy; ACI, adjuvant chemo-immunotherapy; BiTP, anti-PD-L1/TGF-β antibody; CTH, chemotherapy; NAC, neo-adjuvant chemotherapy; NACI, neo-adjuvant chemo-immunotherapy; NAT, neo-adjuvant therapy; SRG, surgery.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: Comparison, Standard Deviation, Adjuvant

Journal: International Journal of Surgery (London, England)

Article Title: Single dual-specific anti-PD-L1/TGF-β antibody synergizes with chemotherapy as neoadjuvant treatment for pancreatic ductal adenocarcinoma: a preclinical experimental study

doi: 10.1097/JS9.0000000000001226

Figure Lengend Snippet: BiTP reverses TGF-β-induced EMT (epithelial–mesenchymal transition) of PDAC (pancreatic ductal adenocarcinoma) cells. (A) CCK-8 (Cell Counting Kit-8) assay to measure the effect of BiTP on TGF-β-mediated chemoresistance in PDAC cells. After 10 ng/ml TGF-β1, 10 5 pM BiTP or hIgG treatment for 24 h. HPDE6-C7, CFPAC-1, ASPC-1, and KPC cells growing in 96-well plates were exposed to serial dilutions of gemcitabine and nab-paclitaxel for 72 h and CCK-8 assays was performed. (B) The representative images of IHC (immunohistochemistry) staining of Ki67 in orthotopic PDAC mice model and statistical graph of the percentage of Ki67-positive cells. (C) The representative images of TUNEL staining for apoptosis (green) and nuclei (DAPI, blue) in orthotopic PDAC mice model and statistical graph of the percentage of apoptosis cells. (D) Transwell assays to measure the effect of BiTP on TGF-β-mediated migration and in PDAC cells. After 10 ng/ml TGF-β1, 10 5 pM BiTP or hIgG treatment for 96 h. 1×10 4 CFPAC-1 and KPC cells were seeded in the upper chambers and Transwell assays were performed. (E) Western blotting assays exploring the chemotherapy-induced EMT and the blocking effect of BiTP. (F) The representative images of IHC staining of E-cadherin in orthotopic PDAC mice model. Statistical graph of the AOD (average optical density) of E-cadherin. (G) The representative images of IF staining of Vimentin (green) and α-SMA (rose red) in orthotopic PDAC mice model. Statistical graph of the arbitrary units of Vimentin and α-SMA. (H) The representative images of Masson staining in orthotopic PDAC mice model. Statistical graph of the percentage of collagen volume fraction (%). Data presented in the graphs represent mean±standard deviation (SD). (**** P <0.0001; *** P <0.001; ** P <0.01; * P <0.05; ns P >0.05). BiTP, anti-PD-L1/TGF-β antibody; CTH, chemotherapy; KPC, LSL-Kras(+/G12D);LSL-Trp53(+/R172H);Pdx1-Cre; PDAC, pancreatic ductal adenocarcinoma; PD-L1, programmed cell death 1 ligand 1; TGF-β, transforming growth factor-β.

Article Snippet: Human PDAC cell lines ASPC-1 (with Kras G12D mutation) and CFPAC-1 (with Kras G12V mutation) and human pancreatic ductal epithelial cell line HPDE6-C7 were obtained from China Center for the Conservation of Typical Cultures (CCTCC).

Techniques: CCK-8 Assay, Cell Counting, Immunohistochemistry, Staining, TUNEL Assay, Migration, Western Blot, Blocking Assay, Standard Deviation