scholarly journals Combination of rAd-p53 in situ gene therapy and anti-PD-1 antibody immunotherapy induced anti-tumor activity in mouse syngeneic urogenital cancer models

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Naoto Kunimura ◽  
Koichi Kitagawa ◽  
Ryota Sako ◽  
Keita Narikiyo ◽  
Shoko Tominaga ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Combination Therapy ◽  
In Vivo Experiments ◽  
Cancer Models ◽  
C3h Mice ◽  
Antibody Immunotherapy ◽  
Adenovirus Receptor ◽  
Urogenital Cancers

Abstract In this study we undertook a novel combination therapy using rAd-p53 in situ gene therapy and immunotherapy with immune checkpoint inhibitor (ICI) anti-PD-1 antibody for urogenital cancers. Three mouse syngeneic tumor cell lines, TRAMP-C2 (prostate cancer derived from C57BL/6 mice), MBT-2 (bladder cancer derived from C3H mice) and Renca (kidney cancer derived from BALB/c mice) were used in this study. The highest coxsackie and adenovirus receptor (CAR) mRNA expression was observed in TRAMP-C2 cells, followed by Renca and then MBT-2 cells. Consistent with the CAR expressions, rAd-p53 at 160 multiplicity of infection (MOI) significantly inhibited the cell proliferation of TRAMP-C2 and Renca cells, but not MBT-2 cells. In in vivo experiments, the combination of intratumoral injections of rAd-p53 (1 × 109 plaque-forming units) every other day and intraperitoneal injections of anti-mouse PD-1 antibody (200 μg) twice a week suppressed tumor growth and prolonged survival compared to rAd-p53 or anti-PD-1 antibody monotherapy in both the TRAMP-C2 and Renca models. Our results encourage the clinical development of combination therapy comprised of in situ gene therapy with rAd-p53 and immunotherapy with an ICI anti-PD-1 antibody for urogenital cancers.

scholarly journals Enlisting the Ixodes scapularis Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick—Pathogen Interactions

Pathogens ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 70
Author(s):  
Lourdes Mateos-Hernández ◽  
Natália Pipová ◽  
Eléonore Allain ◽  
Céline Henry ◽  
Clotilde Rouxel ◽  
...  
Keyword(s):  
Cell Line ◽  
Peripheral Nerves ◽  
Ixodes Scapularis ◽  
Embryonic Cell ◽  
Cell Subpopulation ◽  
In Vivo Experiments

Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like Ixodes scapularis embryonic cell line, ISE6, is an effective tool frequently used for examining tick–pathogen interactions. We detected 37 neuropeptide transcripts in the I. scapularis ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, Anaplasma phagocytophilum. The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in A. phagocytophilum-infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of I. scapularis synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete Ixodes neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick–pathogen interactions.

Phosphofructokinase activity and acidosis during short-term tetanic contractions

1991 ◽  
Vol 69 (2) ◽  
pp. 298-304 ◽  
Author(s):  
Lawrence L. Spriet
Keyword(s):  
Glycolytic Enzyme ◽  
Test Tube ◽  
Mammalian Skeletal Muscle ◽  
Short Term ◽  
Short Supply ◽  
In Vivo Experiments ◽  
Anaerobic Energy

Anaerobic energy production is essential for the production of muscular tension when the demand for energy is greater than can be provided aerobically and when oxygen is in short supply. The largest source of anaerobic energy is from the glycolytic pathway. With sustained tetanic contractions, muscle glycolytic activity is high and hydrogen ions (H+) accumulate while tension production decreases. The increasing [H+] and decreasing tension led to the suggestion that H+ inhibits the activity of the regulatory glycolytic enzyme phosphofructokinase (PFK). Early in vitro work confirmed the H+ sensitivity of PFK in the test tube, indicating that little PFK activity should persist at a pH of 6.9–7.0. However, in situ and in vivo experiments suggested that significant PFK activity was maintained during intense contractions when muscle pH decreased to 6.4–6.6. There are several concerns associated with the application of in vitro findings to in vivo exercise situations: (i) there is little in vitro work in mammalian skeletal muscle with substrate and modulator concentrations representative of exercise, (ii) most in vitro analyses of PFK activity are performed following the dilution of the enzyme in mediums with low protein concentration, and (iii) do the modulators identified in vitro exist in high enough in vivo concentrations at rest and during exercise to contribute to the regulation of PFK? More recent in vitro and in situ PFK experiments have overcome some of these concerns. They confirm that during intense, short-term tetanic contractions, PFK activity is well matched to the ATP demand despite decreases in pH to ~6.4–6.5. A combination of decreased inhibitor (ATP) and increased substrate (fructose 6-phosphate) contents coupled with increases in the contents of several positive modulators may be responsible for the maintained PFK activity. This combination reduces the pH-dependent ATP inhibition of PFK and extends the physiological pH range of the enzyme to the range normally measured during this type of muscular activity.Key words: glycolysis, phosphofructokinase, anaerobic metabolism, acidosis.

Antegrade in situ fenestration of aortic stent graft: in-vivo experiments using a pig model

2012 ◽  
Vol 53 (7) ◽  
pp. 754-758 ◽  
Author(s):  
Petri Saari ◽  
Markku Lähteenvuo ◽  
Krista Honkonen ◽  
Hannu Manninen
Keyword(s):  
Stent Graft ◽  
Pig Model ◽  
In Vivo Experiments ◽  
Aortic Stent Graft

scholarly journals An IL-2-grafted antibody immunotherapy with potent efficacy against metastatic cancer

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Dilara Sahin ◽  
Natalia Arenas-Ramirez ◽  
Matthias Rath ◽  
Ufuk Karakus ◽  
Monika Hümbelin ◽  
...  
Keyword(s):  
Single Molecule ◽  
First Generation ◽  
Metastatic Cancer ◽  
Interleukin 2 ◽  
Antigen Binding ◽  
Cancer Models ◽  
Advanced Malignancies ◽  
Antibody Immunotherapy ◽  
Binding Groove

AbstractModified interleukin-2 (IL-2) formulations are being tested in cancer patients. However, IL-2 immunotherapy damages IL-2 receptor (IL-2R)-positive endothelial cells and stimulates IL-2Rα (CD25)-expressing lymphocytes that curtail anti-tumor responses. A first generation of IL-2Rβ (CD122)-biased IL-2s addressed some of these drawbacks. Here, we present a second-generation CD122-biased IL-2, developed by splitting and permanently grafting unmutated human IL-2 (hIL-2) to its antigen-binding groove on the anti-hIL-2 monoclonal antibody NARA1, thereby generating NARA1leukin. In comparison to hIL-2/NARA1 complexes, NARA1leukin shows a longer in vivo half-life, completely avoids association with CD25, and more potently stimulates CD8+ T and natural killer cells. These effects result in strong anti-tumor responses in various pre-clinical cancer models, whereby NARA1leukin consistently surpasses the efficacy of hIL-2/NARA1 complexes in controlling metastatic disease. Collectively, NARA1leukin is a CD122-biased single-molecule construct based on unmutated hIL-2 with potent efficacy against advanced malignancies.

A phase I/II study of adenovirus-mediated reduced expression in immortalized cells (REIC/DKK-3) gene therapy for prostate cancer: An interim report.

2012 ◽  
Vol 30 (5_suppl) ◽  
pp. 161-161
Author(s):  
Katsumi Sasaki ◽  
Yasutomo Nasu ◽  
Masami Watanabe ◽  
Haruki Kaku ◽  
Ryuta Tanimoto ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Gene Therapy ◽  
Phase I ◽  
Malignant Tumors ◽  
Adenovirus Vector ◽  
Metastatic Lesion ◽  
University Hospital ◽  
Tunel Staining ◽  
In Vivo Experiments

161 Background: A new cancer therapeutic gene; REIC/Dkk-3 was found in Okayama university in 2000. In vivo experiments have demonstrated outstanding cancer-selective apoptosis effects and anti-cancer immunoactivity against various types of malignant tumors. With these positive experimental results, a phase I/II study of adenovirus-mediated REIC/Dkk-3 gene therapy for prostate cancer was initiated from January 2011 in Okayama university hospital. Methods: Two major inclusion criteria category were set up; A: A recurrence of prostate cancer following definitive endocrine therapy with/without metastasis, B: A localized prostate cancer which was considered high risk of recurrences following radical surgery according to a nomogram reported by Kattan et al. Replication-defective adenovirus vector expressing REIC/Dkk-3 (Ad-REIC) was injected directly into the prostate (or metastatic lesion in patients who received prostatectomy) in escalating doses from 1.0×1010 to 1.0×1012 viral particle (vp). Each patient in category A received total of 2 times viral injections every 4 weeks. Patients in category B also received total of 2 times viral injections every 2 weeks, then received radical prostatectomy 6 weeks after second viral injection. Results: Three patients in category A, and 9 patients in category B were entered the present study. No significant side effects were observed. In category B, signs of immunological responses in surgically removed specimens (apoptosis induction of cancer cells in TUNEL staining and infiltration of CD8 positive cells (CTL) in immunohistochemical staining) were observed in second dose. These responses were remarkably observed in the third dose. Conclusions: This initial and preliminary report suggests the favorable safety profile and positive clinical responses in a clinical trial of Ad-REIC gene therapy for prostate cancer.

scholarly journals Application of a simple device for "attracting" and "trapping" glioma cells in situ

2021 ◽  
Author(s):  
Haimin Song ◽  
Runwei Yang ◽  
Runbin Lai ◽  
Kaishu Li ◽  
Bowen Ni ◽  
...  
Keyword(s):  
Growth Factor ◽  
Tumor Cells ◽  
Adult Brain ◽  
Tumor Bed ◽  
In Vivo Experiments ◽  
New Strategy ◽  
New Treatment

Glioblastoma multiforme (GBM) is the most malignant adult brain tumor. The current adjuvant therapies for GBM are disappointing, which are based on cytotoxicity strategy. Thus, other ways should be explored to improve the curative effect. According to the strong invasive ability of GBM cells, we assume a new treatment strategy for GBM by developing a new cell trap device (CTD) with some kind of "attractive" medium loaded in it to attract and capture the tumor cells. The in vitro experiment showed that Hepatocyte Growth Factor（HGF）presented stronger chemotaxis on C6 and U87 cell line than the Epidermal Growth Factor (EGF) and Fibroblast Growth Factor (FGF). A simple in vitro CTD loaded with HGF was made and in vivo experiments results showed that HGF successfully attracted tumor cells from tumor bed in situ into the CTD. This study proposes the new strategy for GBM treatment of "attract and trap" tumor cells is proved to be feasible.

Ion-pair strategy for enabling amifostine oral absorption: Rat in situ and in vivo experiments

2013 ◽  
Vol 49 (4) ◽  
pp. 499-504 ◽  
Author(s):  
N. Samiei ◽  
V. Mangas-Sanjuan ◽  
I. González-Álvarez ◽  
M. Foroutan ◽  
A. Shafaati ◽  
...  
Keyword(s):  
Oral Absorption ◽  
Ion Pair ◽  
In Vivo Experiments

Computational modelling of maternal interactions with spermatozoa: potentials and prospects

2011 ◽  
Vol 23 (8) ◽  
pp. 976 ◽  
Author(s):  
Mark Burkitt ◽  
Dawn Walker ◽  
Daniela M. Romano ◽  
Alireza Fazeli
Keyword(s):  
Reproductive Tract ◽  
Computational Modelling ◽  
Female Reproductive Tract ◽  
In Vivo Experiments ◽  
Complex Interactions ◽  
Maternal Communication ◽  
In Silico Models

Understanding the complex interactions between gametes, embryos and the maternal tract is required knowledge for combating infertility and developing new methods of contraception. Here we present some main aspects of spermatozoa interactions with the mammalian oviduct before fertilisation and discuss how computational modelling can be used as an invaluable aid to experimental investigation in this field. A complete predictive computational model of gamete and embryo interactions with the female reproductive tract is a long way off. However, the enormity of this task should not discourage us from working towards it. Computational modelling allows us to investigate aspects of maternal communication with gametes and embryos, which are financially, ethically or practically difficult to look at experimentally. In silico models of maternal communication with gametes and embryos can be used as tools to complement in vivo experiments, in the same way as in vitro and in situ models.

Molecular Biological Aspects of Fibrinolysis

1979 ◽  
Author(s):  
D. Collen
Keyword(s):  
Plasminogen Activator ◽  
Cell Transformation ◽  
Embryo Implantation ◽  
Tissue Destruction ◽  
In Vivo Experiments ◽  
Biological Aspects ◽  
Plasmin Inhibitor

The fibrinolytic enzyme system plays a role in the removal of fibrin from the blood vessels or urinary tract, and also in tissue repair (angiogenesis), cell transformation, macrophage function, ovulation and embryo implantation. Growing endothelial cells, malignant cells, macrophages, granulosa cells and trophoblast cells.produce plasminogen activators which activate plasminogen in the blood or interstitial fluids. Local plasmin formation appears to be a generalized mechanism involved in tissue destruction and repair. Fibrinolysis in the blood seems to be regulated by specific molecular interactions between (tissue) plasminogen activator, plasmin(ogen), fibrin and α2-antiplasmin, the physiological plasmin inhibitor. Plasmin(ogen) contains structures - lysine-binding sites - which mediate its interaction with fibrin and with α2-antiplasmin. When fibrin forms in plasma a small amount of plasminogen is bound via these structures. Plasminogen activator present or released in the blood is strongly adsorbed to the fibrin and activates bound plasminogen in situ. The formed plasmin, which remains transiently complexed to fibrin, both by its lysine-binding site(s) and active center, is only slowly inactivated by α2-antiplasmin, but plasmin which is released from digested fibrin is rapidly and irreversibly neutralized by α2-antiplasmin. Many in vitro and in vivo experiments support this hypothesis for the mechanism of fibrinolysis (Nature 272, 549,1978). Which allows speculation on more efficient therapeutic schemes for thrombolysis.

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