Exosome-based immunotherapies in ovarian cancer development of preclinical models and therapeutics

2021 ◽  
Author(s):  
◽  
Simone Pisano
Keyword(s):  
Ovarian Cancer ◽  
Immune Cell ◽  
Early Stage ◽  
Advanced Ovarian Cancer ◽  
Cancer Therapeutics ◽  
Multicellular Spheroids ◽  
Early Stage Disease ◽  
Main Research

Treatment of ovarian cancer (OC) continues to present clinical challenges. It remains difficult to diagnose early stage disease, and because of this generally presents as advanced stage, with limited therapeutic options and often the development of resistance to chemotherapy. Furthermore, OC is considered as an immunologically “cold” tumour based on the poor response to immunotherapies. This lack of response to immunotherapy remains poorly understood, hence better models are needed in order to elucidate the fundamental mechanisms behind tumour immunosuppression, and for new and innovative therapeutic strategies to be tested. Exosomes have recently emerged as crucial players in the cell-cell communication and content exchange within the tumour microenvironment and have also been investigated for their capacity to act as cancer therapeutics or to be re-engineered to increase their performances and effects. In this work, two main research areas were explored. The first one focused on advanced ovarian cancer modelling. Indeed, the in vitro formation of multicellular spheroids that included a mixture of cancer cells and different macrophage phenotypes was optimised, and their characteristics explored. Additionally, an in vivo advanced OC model was created from immunocompetent mice and the tumours were characterised for their immune infiltrates presence. The ascitic fluid that arose from the tumour-bearing mice was also comprehensively analysed for its immune cells content by mass cytometry for the first time in an ovarian cancer setting. The second part of the work involved the exploration of two innovative OC therapeutics. Dendritic-cell (DC) derived exosomes were obtained from tumour antigen-pulsed DCs and tested for their efficacy both in vitro and in vivo. A semi-synthetic exosomes approach was also tested by forcing monocytic cells through pores of different sizes and obtaining Immune (Cell) Derived Exosome Mimetics (IDEM). IDEM were characterised and tested in vitro on both 2D and spheroid systems.

The relationship between tumor size and stage in early versus advanced ovarian cancer: A retrospective chart review

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 5580-5580
Author(s):  
L. E. Horvath ◽  
T. Werner ◽  
K. Jones
Keyword(s):  
Ovarian Cancer ◽  
Chart Review ◽  
Advanced Disease ◽  
Early Stage ◽  
Abdominal Cavity ◽  
Advanced Ovarian Cancer ◽  
Retrospective Chart Review ◽  
Advanced Stage ◽  
Early Stage Disease ◽  
Stage Disease

5580 Background: Ovarian cancer has a different prognosis between early (I and II) and advanced stage (III and IV). The mechanism of disease progression is unknown, but patients with advanced disease may have a higher propensity for seeding of the abdominal cavity early in the disease process than those with early stage. Theoretically if this is so, then patients with advanced stage should have smaller sized tumors than patients with early stage. Methods: This was a retrospective chart review of patients in the tumor registry in 2003 to 2006. Patients had epithelial ovarian cancer, other cell types were excluded. Only cases with documentation of surgical and pathologic staging and measured dimensions on pathologic specimen were included. Patient stage and all available dimensions measured on diseased ovaries were recorded. The dimensions for each patient were averaged into a single dimension for that patient, and then these measurements were totaled and averaged. Results: There were 110 patients analyzed: 85 with advanced disease, 25 with early stage. The average measurement was 4.8 cm in advanced disease, and was 10.7 cm in early stage disease. This difference was statistically significant (p < 0.001). Conclusions: Overall, patients with early stage ovarian cancer have diseased ovaries that are more than twice as large as those found in advanced disease. This finding supports the fact that early versus advanced ovarian cancer are 2 separate disease processes. Early stage grows locally and does not disseminate, and advanced stage disseminates while the tumor is still relatively small. Theoretically there may be a factor that separates these 2 into different diseases, where advanced disease patients have a substance produced by their tumor that allows for early dissemination, and early stage lacks this substance and only grows locally. Basic science research comparing the tissue microarrays of early versus advanced stage disease may be able to identify this difference. If the difference is found, perhaps therapy can be targeted against this difference. No significant financial relationships to disclose.

scholarly journals Fibroblast membrane-camouflaged nanoparticles for inflammation treatment in the early stage

2021 ◽  
Vol 13 (1) ◽  
Author(s):  
Lizhong Sun ◽  
Libang He ◽  
Wei Wu ◽  
Li Luo ◽  
Mingyue Han ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Immune Cell ◽  
Early Stage ◽  
Specific Treatment ◽  
Cell Types ◽  
Loss Of Function ◽  
Tissue Repair And Regeneration ◽  
Inflammatory Conditions

AbstractUnrestrained inflammation is harmful to tissue repair and regeneration. Immune cell membrane-camouflaged nanoparticles have been proven to show promise as inflammation targets and multitargeted inflammation controls in the treatment of severe inflammation. Prevention and early intervention of inflammation can reduce the risk of irreversible tissue damage and loss of function, but no cell membrane-camouflaged nanotechnology has been reported to achieve stage-specific treatment in these conditions. In this study, we investigated the prophylactic and therapeutic efficacy of fibroblast membrane-camouflaged nanoparticles for topical treatment of early inflammation (early pulpitis as the model) with the help of in-depth bioinformatics and molecular biology investigations in vitro and in vivo. Nanoparticles have been proven to act as sentinels to detect and competitively neutralize invasive Escherichia coli lipopolysaccharide (E. coli LPS) with resident fibroblasts to effectively inhibit the activation of intricate signaling pathways. Moreover, nanoparticles can alleviate the secretion of multiple inflammatory cytokines to achieve multitargeted anti-inflammatory effects, attenuating inflammatory conditions in the early stage. Our work verified the feasibility of fibroblast membrane-camouflaged nanoparticles for inflammation treatment in the early stage, which widens the potential cell types for inflammation regulation.

scholarly journals An Omentum-inspired 3D PEG Hydrogel for Identifying ECM-drivers of Drug Resistant Ovarian Cancer

2019 ◽  
Author(s):  
Elizabeth A. Brooks ◽  
Maria F. Gencoglu ◽  
Daniel C. Corbett ◽  
Kelly R. Stevens ◽  
Shelly R. Peyton
Keyword(s):  
Ovarian Cancer ◽  
Drug Testing ◽  
Drug Response ◽  
Early Stage ◽  
Treatment Options ◽  
Tissue Culture Polystyrene ◽  
Pathology Reports ◽  
Additional Therapy

AbstractOvarian cancer (OvCa) is a challenging disease to treat due to poor screening techniques and late diagnosis. There is an urgent need for additional therapy options, as patients recur in 70% of cases. The limited availability of clinical treatment options could be a result of poor predictions in early stage drug screens on standard tissue culture polystyrene (TCPS). TCPS does not capture the mechanical and biochemical cues that cells experiencein vivo, which can impact how cells will respond to a drug. Therefore, anin vitromodel that captures some of the microenvironment features that the cells experiencein vivocould provide better insights into drug response. In this study, we formed 3D multicellular tumor spheroids (MCTS) in microwells, and encapsulated them in 3D omentum-inspired hydrogels. SKOV-3 MCTS were resistant to Paclitaxel in our 3D hydrogels compared to a monolayer on TCPS. Toward clinical application, we tested cells from patients (ovarian carcinoma ascites spheroids (OCAS)), and drug responses predicted by using the 3D omentum-inspired hydrogels correlated with the reported pathology reports of those same patients. Additionally, we observed the presence of collagen production around the encapsulated SKOV-3 MCTS, but not on TCPS. Our results demonstrated that our 3D omentum-inspired hydrogel is an improvedin vitrodrug testing platform to study OvCa drug response for patient-derived cells, and helped us identify collagen 3 as a potential driver of Paclitaxel resistance.

GENOMIC AND TRANSCRIPTOME PROFILING OF EPITHELIAL OVARIAN CANCER

2008 ◽  
Vol 31 (4) ◽  
pp. 17
Author(s):  
Rebecca JZ Menzies ◽  
Yury V Bukhman ◽  
Nancy F Ng ◽  
Patricia A Shaw ◽  
Tak W Mak
Keyword(s):  
Ovarian Cancer ◽  
Epithelial Ovarian Cancer ◽  
Early Stage ◽  
Snp Array ◽  
Ovarian Tumors ◽  
Ca 125 ◽  
Molecular Heterogeneity ◽  
Presenting Symptoms

Background: Epithelial ovarian cancer is the leading cause of death by gynecological malignancy. Due to inadequate screening modalities, a lack of characteristic presenting symptoms, limited treatments and a poor understanding of the molecular underpinnings of the disease, only 25% of ovarian cancers are diagnosed at an early stage. Current 5-year survival rates range from 80%, for disease diagnosed in Stage I to as low as 13% for Stage IV. Current screening for ovarian cancer involves measuring CA-125 levels. However, CA-125 testing has low sensitivity since it can be elevated in a variety of other gynecological diseases. Numerous studies have found molecular heterogeneity between the four histological subtypes of ovarian cancer (serous, endometrioid, clear cell and mucinous). However, treatments remain the same for all subtypes regardless of molecular heterogeneity. Thus, better treatment targets and biomarkers must be found for this disease. Methods:In our study 300 ovarian tumors will be genomically profiled using the Affymetrix Genome-Wide SNP Array 6.0 to identify loci and genes implicated in ovarian cancer. To date, 51 ovarian tumors have been analyzed using the SNP array. Results:Preliminary analysis of copy number variation in these tumors using Partek software has revealed a total of 978 loci. Known amplifications derived from the literature were seen at CCNE1 and ERBB2. Similarly, well known deletions ofp53 and RB1 in ovarian cancer were detected. Novel amplified loci at 18q11.2 and 4q33 were also detected. Novel deletions were detected at 7p13 and 8q22.2. Conclusion: Future work will include running the remaining 249 tumor samples on the SNP array and analyzing the complete dataset using Partek software. Future validation of identified genes in vitro and in vivo may provide insight and possible biomarkers that may be used clinically to benefit the ovarian cancer patient.

scholarly journals Regenerative Medicine and Angiogenesis; Focused on Cardiovascular Disease

2021 ◽  
Author(s):  
Seyed Zachariah Moradi ◽  
Faramarz Jalili ◽  
Zohreh Hoseinkhani ◽  
Kamran Mansouri
Keyword(s):  
Cardiovascular Disease ◽  
Early Stage ◽  
In Vitro Models ◽  
Early Stage Disease ◽  
Surgical Interventions ◽  
Beneficial Effects ◽  
Regeneration Medicine ◽  
Affected Tissue

Cardiovascular disease (CVD) is a major concern for health with high mortality rates around the world. CVD is often associated with partial or full occlusion of the blood vessel network. Changes in lifestyle can be useful for management early-stage disease but in the advanced stage, surgical interventions or pharmacological are needed to increase the blood flow through the affected tissue or to reduce the energy requirements. Regeneration medicine is a new science that has provided many different options for treating various diseases, especially in CVD over the years. Stem cell therapy, gene therapy, and tissue engineering are some of the powerful branches of the field that have given patients great hope in improving their condition. In this review, we attempted to examine the beneficial effects, challenges, and contradictory effects of angiogenesis in vivo, and in vitro models’ studies of CVD. We hope that this information will be able to help other researchers to design new effective structures and open new avenues for the treatment of CVD with the help of angiogenesis and regeneration medicine in the future.

scholarly journals Endangered Lymphocytes: The Effects of Alloxan and Streptozotocin on Immune Cells in Type 1 Induced Diabetes

2021 ◽  
Vol 2021 ◽  
pp. 1-15
Author(s):  
Luiz A. D. Queiroz ◽  
Josiane B. Assis ◽  
João P. T. Guimarães ◽  
Emanuella S. A. Sousa ◽  
Anália C. Milhomem ◽  
...  
Keyword(s):  
Immune Cell ◽  
Early Stage ◽  
Chronic Phase ◽  
Treated Group ◽  
Inflammatory Infiltration ◽  
Cell Parameters ◽  
Pancreatic Islets Of Langerhans

Alloxan (ALX) and streptozotocin (STZ) are extensively used to induce type 1 diabetes (T1D) in animal models. This study is aimed at evaluating the differences in immune parameters caused by ALX and STZ. T1D was induced either with ALX or with STZ, and the animals were followed for up to 180 days. Both ALX and STZ induced a decrease in the total number of circulating leukocytes and lymphocytes, with an increase in granulocytes when compared to control mice (CT). STZ-treated mice also exhibited an increase in neutrophils and a reduction in the lymphocyte percentage in the bone marrow. In addition, while the STZ-treated group showed a decrease in total CD3+, CD4-CD8+, and CD4+CD8+ T lymphocytes in the thymus and CD19+ B lymphocytes in the pancreas and spleen, the ALX group showed an increase in CD4-CD8+ and CD19+ only in the thymus. Basal levels of splenic interleukin- (IL-) 1β and pancreatic IL-6 in the STZ group were decreased. Both diabetic groups showed atrophy of the thymic medulla and degeneration of pancreatic islets of Langerhans composed of inflammatory infiltration and hyperemia with vasodilation. ALX-treated mice showed a decrease in reticuloendothelial cells, enhanced lymphocyte/thymocyte cell death, and increased number of Hassall’s corpuscles. Reduced in vitro activation of splenic lymphocytes was found in the STZ-treated group. Furthermore, mice immunized with ovalbumin (OVA) showed a more intense antigen-specific paw edema response in the STZ-treated group, while production of anti-OVA IgG1 antibodies was similar in both groups. Thereby, important changes in immune cell parameters in vivo and in vitro were found at an early stage of T1D in the STZ-treated group, whereas alterations in the ALX-treated group were mostly found in the chronic phase of T1D, including increased mortality rates. These findings suggest that the effects of ALX and STZ influenced, at different times, lymphoid organs and their cell populations.

scholarly journals GZMKhigh CD8+ T effector memory cells are associated with CD15high neutrophil abundance in early-stage colorectal tumors and predict poor clinical outcome.

2021 ◽  
Author(s):  
Silvia Tiberti ◽  
Carlotta Catozzi ◽  
Caterina Scirgolea ◽  
Ottavio Croci ◽  
Mattia Ballerini ◽  
...  
Keyword(s):  
T Cells ◽  
Clinical Outcome ◽  
Cd8 T Cells ◽  
Immune Cell ◽  
Early Stage ◽  
Effector Memory ◽  
Functional Changes

Tumor contexture has emerged as a major determinant to establish prognosis and guide novel therapies and tumor infiltrating CD8+ T cells have been associated with a better prognosis in several solid tumors, including early-stage colorectal cancer (CRC). However, the tumor immune infiltrate is highly heterogeneous and understanding how the interplay between different immune cell compartments impacts on the clinical outcome is still in its infancy. Here, we describe in a prospective cohort a novel CD8+ T effector memory population, which is characterized by high levels of Granzyme K (GZMKhigh CD8+ TEM) and correlated with CD15high tumor infiltrating neutrophils. We provide both in vitro and in vivo evidence of the role of stromal cell-derived factor 1 (CXCL12/SDF-1) in driving functional changes on neutrophils at the tumor site, promoting their retention and increasing crosstalk with CD8+ T cells. Mechanistically, as a consequence of the interaction with neutrophils, CD8+ T cells are skewed towards a CD8+ TEM phenotype, producing high levels of GZMK, which in turn decreased E-cadherin pathway. The correlation of GZMKhigh CD8+ TEM and neutrophils with both tumor progression in mice and early relapse in CRC patients demonstrate the role of GZMKhigh CD8+ TEM in promoting malignancy. Indeed, a gene signature defining GZMKhigh CD8+ TEM was associated with worse prognosis on a larger independent cohort of CRC patients and a similar analysis was extended to lung cancer (TCGA). Overall, our results highlight the emergence of GZMKhigh CD8+ TEM in early-stage CRC tumors as a hallmark driven by the interaction with neutrophils, which could implement current patient stratification and be targeted by novel therapeutics.

707 Discovery of a novel EP2 and EP4 dual antagonist

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A749-A749
Author(s):  
Yeri Lee ◽  
Hyun Jin Kim ◽  
Donggeon Kim ◽  
Joo Youn Lee ◽  
Chang Soo Yun ◽  
...  
Keyword(s):  
Cell Function ◽  
Immune Cell ◽  
Radioligand Binding ◽  
Early Stage ◽  
Scintillation Counter ◽  
Hek293 Cells ◽  
List Type ◽  
Camp Levels

BackgroundProstaglandin E2 (PGE2) is one of the most abundant prostaglandins, with crucial roles in normal and pathologic physiology. Especially, PGE2 levels are abnormally elevated in many cancers, and high levels of PGE2 are known to be pro-tumorigenic, likely due to the immune suppressive effect in the tumor microenvironment.1–4 There are four types of PGE2 receptors; EP1, EP2, EP3 and EP4. Among them, EP2 and EP4 activate adenylate cyclase and increase cAMP levels, which induce the cAMP-dependent protein kinase (PKA) signaling pathway. EP2 and EP4 are expressed in various immune cells (e.g. macrophages, dendritic cells, NK cells and CTLs), and genetic and pharmacological inhibition of EP2 and EP4 increases immune activity and suppresses tumor growth.MethodsTo evaluate the binding affinity against EP2 and EP4, a radioligand binding assay was conducted using EP2 or EP4 transfected HEK293 cells. Cell membrane homogenates were incubated with [3H]PGE2 in the absence or presence of the test compounds. Following incubation, the samples were filtered rapidly under vacuum through glass fiber filters and rinsed several times with cold Tris-HCl. The filters were dried then counted for radioactivity in a scintillation counter using a scintillation cocktail. The results were expressed as a percent inhibition of the control radioligand specific binding.The antagonistic activity against EP2 and EP4 was assessed via LANCE Ultra cAMP assay (PerkinElmer). HEK293 cells overexpressing EP2 or EP4 were seeded into the plate and treated by PGE2 and compounds. After 30 minutes of incubation, cAMP levels were measured by FRET signal using Varioskan plate reader, following the manufacturer’s protocol.Anti-tumor activity of KT-00113 was evaluated using LLC1 syngeneic model. When tumor volume reached approximatively 100 mm3, mice were treated PO, QD. Tumor size was measured twice every week.ResultsSystematic structure-activity relationship (SAR) investigation identified novel EP2 and EP4 dual antagonists. The most promising compound KT-00113 possesses high potency against both EP2 and EP4, while maintaining high selectivity over other prostanoid receptors. In vitro and in vivo ADMET studies show that KT-00113 has a favorable profile, apt for further examination in in vivo cancer models and immune cell function in tumors.ConclusionsKT-00113, a highly potent and selective EP2/4 dual antagonist has strong potential to become the best-in-class immune suppression lifting cancer immunotherapy and may be suitable for further development in a clinical setting.ReferencesCordes T, Hoellen F, Dittmer C, Salehin D, Kummel S, Friedrich M, Koster F, Becker S, Diedrich K, Thill M. Correlation of prostaglandin metabolizing enzymes and serum PGE2 levels with vitamin D receptor and serum 25(OH)2D3 levels in breast and ovarian cancer. Anticancer Res;32 ( 2012):351–357.Diakowska D, Markocka-Maczka K, Nienartowicz M, Lewandowski A, Grabowski K. Increased level of serum prostaglandin-2 in early stage of esophageal squamous cell carcinoma, Arch Med Sci ( 2014);10:956–961.Akbari N, Ghorbani M, Salimi V, Alimohammadi A, Khamseh ME, Akbari H, Nourbakhsh M, Sheikhi A, Taghavi SF, Tavakoli-Yaraki M. Cyclooxygenase enzyme and PGE2 expression in patients with functional and non-functional pituitary adenomas. BMC Endocr Disord 2020;20:39.Gomes RN, Felipe da Costa S, Colquhoun A. Eicosanoids and cancer, Clinics (Sao Paulo), 73 ( 2018) e530s.

scholarly journals Emerging Roles for Ion Channels in Ovarian Cancer: Pathomechanisms and Pharmacological Treatment

Cancers ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 668
Author(s):  
Concetta Altamura ◽  
Maria Raffaella Greco ◽  
Maria Rosaria Carratù ◽  
Rosa Angela Cardone ◽  
Jean-François Desaphy
Keyword(s):  
Ovarian Cancer ◽  
Ion Channels ◽  
Transient Receptor Potential ◽  
Critical Role ◽  
Gynecologic Cancer ◽  
Receptor Potential ◽  
Transient Receptor Potential Channels ◽  
Platinum Resistance

Ovarian cancer (OC) is the deadliest gynecologic cancer, due to late diagnosis, development of platinum resistance, and inadequate alternative therapy. It has been demonstrated that membrane ion channels play important roles in cancer processes, including cell proliferation, apoptosis, motility, and invasion. Here, we review the contribution of ion channels in the development and progression of OC, evaluating their potential in clinical management. Increased expression of voltage-gated and epithelial sodium channels has been detected in OC cells and tissues and shown to be involved in cancer proliferation and invasion. Potassium and calcium channels have been found to play a critical role in the control of cell cycle and in the resistance to apoptosis, promoting tumor growth and recurrence. Overexpression of chloride and transient receptor potential channels was found both in vitro and in vivo, supporting their contribution to OC. Furthermore, ion channels have been shown to influence the sensitivity of OC cells to neoplastic drugs, suggesting a critical role in chemotherapy resistance. The study of ion channels expression and function in OC can improve our understanding of pathophysiology and pave the way for identifying ion channels as potential targets for tumor diagnosis and treatment.

Three-dimensional Growth of Renal Epithelial Cells in Vitro: A Tool in Toxicity Testing

1993 ◽  
Vol 21 (2) ◽  
pp. 191-195 ◽  
Author(s):  
Knut-Jan Andersen ◽  
Erik Ilsø Christensen ◽  
Hogne Vik
Keyword(s):  
Epithelial Cells ◽  
Three Dimensional ◽  
Toxicity Testing ◽  
Renal Tissue ◽  
Epithelial Cell Line ◽  
Multicellular Spheroids ◽  
Renal Epithelial Cell ◽  
Renal Epithelial Cells

The tissue culture of multicellular spheroids from the renal epithelial cell line LLC-PK1 (proximal tubule) is described. This represents a biological system of intermediate complexity between renal tissue in vivo and simple monolayer cultures. The multicellular structures, which show many similarities to kidney tubules in vivo, including a vectorial water transport, should prove useful for studying the potential nephrotoxicity of drugs and chemicals in vitro. In addition, the propagation of renal epithelial cells as multicellular spheroids in serum-free culture may provide information on the release of specific biological parameters, which may be suppressed or masked in serum-supplemented media.

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