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IRCT20170208032451N1 IRCT 2020-09-14 Mashhad University of Medical Sciences Gastric cancer immunotherapy Clinical evaluation of immunogenic adjuvant therapy with dendritic cells loaded with autologous tumor mRNA in patients with gastric cancer 2021-01-20 anticipated 5 Complete https://irct.ir/trial/49923 interventional Randomization: N/A, Blinding: Not blinded, Placebo: Not used, Assignment: Single, Purpose: Treatment. 1-2 gastric cancer. Chimer Construction Design: using immunogenetic epitopes of MAGEA4, LAGE1, and NY-ESO1 antigens, a chimeric molecule is prepared which, due to the pivotal role of dendritic cells in inducing an immune response, is sent into these cells in the form of mRNA to stimulate the immune system. Provide gastric cancer patients. Due to the overexpression of selective markers (MAGE-A4, LAGE1, and NY-ESO1) in gastric tumor cells compared to normal cells, the structural basis of the construct molecule was based on specific sequences of the same genes. Our goal is to identify these proteins to lymphocyte cells as tumor markers. Since it is difficult to transfer the complete gene or mRNA of all three markers to the antigen-supplying cells, parts of each protein were selected and synthesized together into one molecule.Construction of contraceptive mRNA by Mmessage Mmachin kit:Plasmid PGEM-4Z / GFP / A64, which has a polymythine sequence at the end of the transcription region, is used as the target vector for the synthesized construct. Chimeric Antigene mRNA amplification is performed using an in vitro transcription reaction.Leukophoresis and isolation of monocytes from peripheral blood:Isolation of diseased monocytes and lymphocytes from peripheral blood mononuclear cells (PBMC) is performed by leukophoresis. After isolation of monocytes and lymphocytes by specific leukophoresis kits, the cells were transferred to the laboratory to be converted to DC cells in vitro. In order to isolate T lymphocytes, which are required in the next stages of the test, using the conventional method of attaching monocyte cells to the bottom of the culture flask, the unattached cells are the same T lymphocytes that are used for the next steps of the test. Placed.Production of dendritic immature cells (DC immature): According to the standard instructions of blood monocytes, under laboratory culture and with the addition of cytokines GM-CSF, IL-4 at specific concentrations within 5 to 7 days, monocytes become immature dendritic cells. At the end of this stage, the produced cells of dendritic cell or DC immature are evaluated by flow cytometry.Transfer of Chimeric Antigene mRNA to immature DC cells (DC loading):At this stage, immature DC cells are transferred to the DC cell with Chimeric Antigene mRNA 25 μg / mL by nanoparticle.DC maturation stage by cytokine mixture:At this stage, the loaded DCs are placed in the vicinity of the maturation stimuli to mature. In RPMI-1640 culture medium, cytokine cocktail containing a mixture of cytokines IL-1, IL-6, PGE2, TNF- is performed in 2-3 days of DC cell maturation stages.Phenotypic study of produced DC cells:This operation is performed by flow cytometry device and monoclonal antibodies by examining CCR7, CD83, CD14, CD86, CD80 and HLA-DR markers.648/5000Vaccination schedule:Patients are divided into two groups based on cell dose levels. The first measurable patient was vaccinated with a low dose (injection / cell dose 107.1), the second measurable patient with a moderate dose (injection / cell dose 107.3), and the third measurable patient with a high dose (injection / cell dose 108.1). To be. Four consecutive vaccinations with dendritic cells loaded with chimeric antigens are performed intravenously once a week (21-day period). The fourth and fifth patients inject dendritic cells loaded with dendritic antigens intravenously (injection / cell dose 106 × 1) intradermally into the forearm or thigh 4 times with an interval of two weeks between each vaccine. To turn.. No - There is not a plan to make this available Justification or reason for not sharing IPD is no more information
public Mohammad Reza Rbbaszadegan
Ferdowsi University
Mashhad Iran (Islamic Republic of) 9177948564 +98 51 3800 2000 abbaszadeganmr@mums.ac.ir Mashhad University of Medical Sciences scientific Mohammad Reza Abbaszadegan
East door of Ferdowsi University, Public Relations Department of the University
Mashhad Iran (Islamic Republic of) 9177899191 0985138002000 abbaszadeganmr@mums.ac.ir Mashhad University of Medical Sciences Iran (Islamic Republic of) Iran (Islamic Republic of) Patients with gastric adenocarcinoma whose histologically advanced malignancy (spread of cancer cells to lymph nodes and tumor metastasis) has been confirmed and they have filled out the informed consent form The patient must be over 18 years old at the time of diagnosis. Natural laboratory parameters, including these tests: Protein C& S, D-Dimer, VWF, CPK,LDH, TI, ALP, Lipase, Amylase, GGT, OB,CBC diff, Hgb electrophoresis, Serum Iron, Transferrin saturation, TIBC, CH50 (total complement), ESR, CRP, ACE, ANA, Anti CCP, RF, Auto antibodies, UA, Uric acid, Cr, Urea, SGPT, SGOT, Bilirubin, Albumin, Lipid profile, ACTH, Cortisol, FBS, HbA1c, OGT , ACTH, Cortisol, FBS, HbA1c, OGTT, Insulin, PTH, GH, Calcitonin, TFT, Testosterone, LH, FSH, Estrogen, Progestrone, PRL. To start immunotherapy, at least 4 weeks must have passed since the last course of chemotherapy or radiotherapy, and all the side effects of chemotherapy must be eliminated. Functional condition according to Karnowski criteria should be more than 60% Expected survival rate more than 6 months 18 years 80 years Both brain metastasis autoimmune diseases Abnormal function of heart, liver, kidney, brain and other important organs C16 Malignant neoplasm of stomach Treatment - Other Chimer Construction Design: using immunogenetic epitopes of MAGEA4, LAGE1, and NY-ESO1 antigens, a chimeric molecule is prepared which, due to the pivotal role of dendritic cells in inducing an immune response, is sent into these cells in the form of mRNA to stimulate the immune system. Provide gastric cancer patients. Due to the overexpression of selective markers (MAGE-A4, LAGE1, and NY-ESO1) in gastric tumor cells compared to normal cells, the structural basis of the construct molecule was based on specific sequences of the same genes. Our goal is to identify these proteins to lymphocyte cells as tumor markers. Since it is difficult to transfer the complete gene or mRNA of all three markers to the antigen-supplying cells, parts of each protein were selected and synthesized together into one molecule.Construction of contraceptive mRNA by Mmessage Mmachin kit:Plasmid PGEM-4Z / GFP / A64, which has a polymythine sequence at the end of the transcription region, is used as the target vector for the synthesized construct. Chimeric Antigene mRNA amplification is performed using an in vitro transcription reaction.Leukophoresis and isolation of monocytes from peripheral blood:Isolation of diseased monocytes and lymphocytes from peripheral blood mononuclear cells (PBMC) is performed by leukophoresis. After isolation of monocytes and lymphocytes by specific leukophoresis kits, the cells were transferred to the laboratory to be converted to DC cells in vitro. In order to isolate T lymphocytes, which are required in the next stages of the test, using the conventional method of attaching monocyte cells to the bottom of the culture flask, the unattached cells are the same T lymphocytes that are used for the next steps of the test. Placed.Production of dendritic immature cells (DC immature): According to the standard instructions of blood monocytes, under laboratory culture and with the addition of cytokines GM-CSF, IL-4 at specific concentrations within 5 to 7 days, monocytes become immature dendritic cells. At the end of this stage, the produced cells of dendritic cell or DC immature are evaluated by flow cytometry.Transfer of Chimeric Antigene mRNA to immature DC cells (DC loading):At this stage, immature DC cells are transferred to the DC cell with Chimeric Antigene mRNA 25 μg / mL by nanoparticle.DC maturation stage by cytokine mixture:At this stage, the loaded DCs are placed in the vicinity of the maturation stimuli to mature. In RPMI-1640 culture medium, cytokine cocktail containing a mixture of cytokines IL-1, IL-6, PGE2, TNF- is performed in 2-3 days of DC cell maturation stages.Phenotypic study of produced DC cells:This operation is performed by flow cytometry device and monoclonal antibodies by examining CCR7, CD83, CD14, CD86, CD80 and HLA-DR markers.648/5000Vaccination schedule:Patients are divided into two groups based on cell dose levels. The first measurable patient was vaccinated with a low dose (injection / cell dose 107.1), the second measurable patient with a moderate dose (injection / cell dose 107.3), and the third measurable patient with a high dose (injection / cell dose 108.1). To be. Four consecutive vaccinations with dendritic cells loaded with chimeric antigens are performed intravenously once a week (21-day period). The fourth and fifth patients inject dendritic cells loaded with dendritic antigens intravenously (injection / cell dose 106 × 1) intradermally into the forearm or thigh 4 times with an interval of two weeks between each vaccine. To turn. Side effect. Timepoint: 7 days. Method of measurement: clinical measurement. Clinical and immune response. Timepoint: one year. Method of measurement: Flow cytometry - ELISA - Overall survival rate - Tumor-free survival rate. Mashhad University of Medical Sciences Approved 2020-01-28 mashhad university of medical sciences East door of Ferdowsi University, Public Relations Department of the University mashhad Razavi Khorasan Iran (Islamic Republic of)