pyrophosphate and Adenocarcinoma

Pancreatic adenocarcinoma (PDAC) is highly refractory to treatment. Standard-of-care gemcitabine (Gem) provides only modest survival benefits, and development of Gem resistance (GemR) compromises its efficacy. Highly GemR clones of Gem-sensitive MIAPaCa-2 cells were developed to investigate the molecular mechanisms of GemR and implemented global quantitative differential proteomics analysis with a comprehensive, reproducible ion-current-based MS1 workflow to quantify ∼6000 proteins in all samples. In GemR clone MIA-GR8, cellular metabolism, proliferation, migration, and 'drug response' mechanisms were the predominant biological processes altered, consistent with cell phenotypic alterations in cell cycle and motility. S100 calcium binding protein A4 was the most downregulated protein, as were proteins associated with glycolytic and oxidative energy production. Both responses would reduce tumor proliferation. Upregulation of mesenchymal markers was prominent, and cellular invasiveness increased. Key enzymes in Gem metabolism pathways were altered such that intracellular utilization of Gem would decrease. Ribonucleoside-diphosphate reductase large subunit was the most elevated Gem metabolizing protein, supporting its critical role in GemR. Lower Ribonucleoside-diphosphate reductase large subunit expression is associated with better clinical outcomes in PDAC, and its downregulation paralleled reduced MIAPaCa-2 proliferation and migration and increased Gem sensitivity. Temporal protein-level Gem responses of MIAPaCa-2 versus GemR cell lines (intrinsically GemR PANC-1 and acquired GemR MIA-GR8) implicate adaptive changes in cellular response systems for cell proliferation and drug transport and metabolism, which reduce cytotoxic Gem metabolites, in DNA repair, and additional responses, as key contributors to the complexity of GemR in PDAC. These findings additionally suggest targetable therapeutic vulnerabilities for GemR PDAC patients.

Topics: Adenocarcinoma; Cell Line, Tumor; Diphosphates; Drug Resistance, Neoplasm; Gemcitabine; Humans; Pancreatic Neoplasms; Proteomics; Ribonucleosides; S100 Calcium-Binding Protein A4

Lysophosphatidic acid (LPA) mediates a variety of cellular responses with atleast six G protein-coupled transmembrane receptors (LPA receptor-1 (LPA(1)-LPA(6))). The interaction between LPA receptors and other cellular molecules on the biological function is not fully understood. Recently, we have reported that LPA(1) suppressed and LPA(3) stimulated cell migration of pancreatic cancer cells. In the present study, to evaluate the function of LPA(2) on motile and invasive activities of pancreatic cancer cells, we generated Lpar2 knockdown (HPD-sh2) cells from hamster pancreatic cancer cells and measured their cell migration ability. In cell motility and invasive assays with an uncoated Cell Culture Insert, HPD-sh2 cells showed significantly lower intrinsic activity than control (HPD-GFP) cells. Since K-ras mutations were frequently detected in pancreatic cancer, we next investigated whether oncogenic K-ras is involved in cell migration induced by LPA(2) using K-ras knockdown (HPD-K2) cells. The cell motile ability of HPD-K2 cells was significantly lower than that of control cells. To confirm LPA(2) increases cell migration activity, cells were pretreated with dioctylglycerol pyrophosphate (DGPP) which is the antagonist of LPA(1)/LPA(3). The cell motile and invasive abilities of DGPP -treated HPD-GFP cells were markedly higher than those of untreated cells, but DGPP did not stimulate cell migration of HPD-K2 cells. These results suggest that cell migration activity of pancreatic cancer cells stimulated by LPA(2) may be enhanced by oncogenic K-ras.

Topics: Adenocarcinoma; Animals; Carcinoma, Pancreatic Ductal; Cell Line, Tumor; Cell Movement; Cricetinae; Diphosphates; Gene Knockdown Techniques; Genes, ras; Glycerol; Lysophospholipids; Neoplasm Invasiveness; Pancreatic Neoplasms; Receptors, Lysophosphatidic Acid; Up-Regulation

Hepatocellular carcinoma (HCC) and colorectal carcinoma with hepatic metastases (mCRC) are cancers with poor prognosis and limited therapeutic options. New approaches are needed and adoptive immunotherapy with Vgamma9Vdelta2 T lymphocytes represents an attractive strategy. Indeed, Vgamma9Vdelta2 T cells were shown to exhibit efficient lytic activity against various human tumor cell lines, and in vitro Vgamma9Vdelta2 T expansion protocol based on single phosphoantigen stimulation could be easily performed for healthy donors. However, a low proliferative response of Vgamma9Vdelta2 T cells was observed in about half of the cancer patients, leading to an important limitation in the development of Vgamma9Vdelta2 T cell-based immunotherapy. Here, for the first time in the context of cancer patients, Vgamma9Vdelta2 T cell expansions were performed by co-culturing peripheral blood mononuclear cell (PBMCs) with autologous dendritic cells (DCs) pretreated with aminobisphosphonate zoledronate. For patients not responding to the conventional culture protocol, co-culture of PBMC with zoledronate-pretreated DCs induced strong cell expansion and allowed reaching a minimal rate of purity of 70% of Vgamma9Vdelta2 T cells. The potent immunostimulatory activity of zoledronate-treated DCs was associated with higher amount of isopentenyl pyrophosphate (IPP) in the culture and was correlated with better ability to activate Vgamma9Vdelta2 T cells as measured by IFN-gamma production. Moreover, we demonstrated that the cytotoxic level of Vgamma9Vdelta2 T cells against freshly autologous tumor cells isolated from patients could be significantly increased by pretreating the tumor cells with zoledronate. Thus, this method of generating Vgamma9Vdelta2 T cells leads eligible for Vgamma9Vdelta2 T cell adoptive immunotherapy the HCC and mCRC patients.

Topics: Adenocarcinoma; Aged; Blotting, Western; Bone Density Conservation Agents; Carcinoma, Hepatocellular; Cell Differentiation; Cell Proliferation; Coculture Techniques; Colorectal Neoplasms; Cytotoxicity, Immunologic; Dendritic Cells; Diphosphates; Diphosphonates; Female; Flow Cytometry; Hemiterpenes; Humans; Imidazoles; Immunotherapy, Adoptive; Liver Neoplasms; Male; Middle Aged; Organophosphorus Compounds; Receptors, Antigen, T-Cell, gamma-delta; T-Lymphocyte Subsets; Zoledronic Acid

Radionuclide bone scans and skeletal radiographs were obtained before and during combination chemotherapy or initial hormonal treatment in 46 patients with disseminated adenocarcinoma of the prostate. The purpose of the study was to determine the usefulness of these two modalities in evaluating tumor response to therapy. Prior to treatment, bone scans were positive in 44 patients (96%). In all but one patient either bone radiographs or bone marrow biopsy revealed evidence of osseous metastases. In 22 patients partial response to therapy was documented by a variety of other staging tests. Eleven of these patients showed concurrent or later improvement on bone scans; one showed improvement on a radiograph. "Flare phenomena" were observed relatively frequently since 23% of the scans and 50% of the radiographs showed worsening at the time of response. Bone scans revealed worsening in 79% of 33 patients with disease progression of extraosseous tumor; radiographs were equally sensitive (82% worsening). It is concluded that bone scans in particular are useful for monitoring tumor status in systemically treated patients with prostate cancer. However, because of the lack of sensitivity for response and paradoxical worsening with tumor regression in some patients, scans are not accurate enough to be employed as the sole test in following these patients.

Topics: Adenocarcinoma; Aged; Bone Neoplasms; Castration; Cisplatin; Cyclophosphamide; Diphosphates; Diphosphonates; Doxorubicin; Drug Therapy, Combination; Humans; Male; Middle Aged; Prostatic Neoplasms; Technetium; Technetium Tc 99m Medronate; Technetium Tc 99m Pyrophosphate

The low incidence of measurable or evaluable metastases in patients with prostatic cancer makes evaluation of response difficult. This is particularly true in patients with bone metastases only. With a digital model it is possible to measure quantitatively from the radioisotope bone scan the total area of skeletal involvement by metastatic tumor. Definitions of response in bone have been derived from this model. These response criteria have been compared to response in acid phosphatase determinations and clinical status in a study of 44 patients with advanced prostatic cancer treated with estramustine phosphate. Based on serial quantitative bone scans, serial measurements of acid phosphatase levels and repeat clinical evaluations a system is proposed for defining response to systemic therapy that is applicable to the majority of patients with metastatic prostatic cancer.

Topics: Acid Phosphatase; Adenocarcinoma; Adult; Aged; Bone Neoplasms; Diphosphates; Diphosphonates; Estramustine; Humans; Male; Middle Aged; Nitrogen Mustard Compounds; Prostatic Neoplasms; Radionuclide Imaging; Technetium

Topics: Adenocarcinoma; Aged; Bone and Bones; Diphosphates; Humans; Kidney Neoplasms; Male; Radionuclide Imaging; Technetium

Topics: Adenocarcinoma; Adenocarcinoma, Scirrhous; Adult; Bone Neoplasms; Breast Neoplasms; Carcinoma; Diphosphates; Female; Humans; Middle Aged; Neoplasm Metastasis; Radionuclide Imaging; Technetium

Bone scan findings (using 99mTc-stannous pyrophosphate) in five patients with diffuse metastatic carcinoma of the axial skeleton are reviewed. Although there were few visually recognizable asymmetries of tracer localization, the diffuse involvement was diagnosed through abnormally elevated counting rates in the axial skeleton, decreased visualization of the kidneys, and faint or absent visualization of the appendicular skeleton.

Topics: Adenocarcinoma; Aged; Bone Neoplasms; Carcinoma, Transitional Cell; Diphosphates; Humans; Kidney Neoplasms; Kidney Pelvis; Male; Middle Aged; Neoplasm Metastasis; Prostatic Neoplasms; Radionuclide Imaging; Ribs; Skull Neoplasms; Spinal Neoplasms; Sternum; Technetium

A study was undertaken to investigate the behavior of 99mTc-Sn-pyrophosphate complex in metabolic bone disease. Of clinical importance was the generalized increased periarticular bone accumulation of the radiopharmaceutical in osteomalacia and in combined osteomalacia and osteitis fibrosa as found in patients with chronic renal failure. The pattern in primary hyperparathyroidism was variable. There was no correlation between the initial rates of accumulation of the radiophosphate complex or its bone to soft-tissue uptake ratio at 5 hr when compared with the degree of osteomalacia and osteitis fibrosa. It is postulated that the 99mTc-Sn-pyrophosphate complex has greater affinity for immature collagen than the crystal surface.

Topics: Adenocarcinoma; Adult; Aged; Anorexia Nervosa; Bone Diseases; Diphosphates; Female; Humans; Hyperparathyroidism; Hyperparathyroidism, Secondary; Middle Aged; Osteitis Deformans; Osteitis Fibrosa Cystica; Osteomalacia; Parathyroid Neoplasms; Radionuclide Imaging; Renal Dialysis; Technetium

A retrospective study of 795 consecutive bone scans employing either 18F or 99mTc-pyrophosphate to evaluate the diagnostic value of renal asymmetry in such scans has been carried out. It is concluded that asymmetric renal images in bone scans convey relatively specific information regarding renal pathology, especially in the 99mTc-pyrophosphate studies.

Topics: Adenocarcinoma; Adult; Aged; Bone Neoplasms; Carcinoma, Bronchogenic; Diphosphates; Female; Fluorine; Humans; Kidney; Kidney Diseases; Kidney Neoplasms; Male; Melanoma; Radioisotopes; Radionuclide Imaging; Retrospective Studies; Technetium; Ureteral Obstruction

Topics: Adenocarcinoma; Azoles; Binding Sites; Carbon Isotopes; Culture Techniques; Diphosphates; Female; Glutamine; Hot Temperature; Hydrogen-Ion Concentration; Kinetics; Magnesium; Mercaptopurine; Neoplasms, Experimental; Nucleotides; Quaternary Ammonium Compounds; Ribose; Sulfates; Transferases