Establishment of In Vivo Ovarian Cancer Mouse Models Using Intraperitoneal Tumor Cell Injection

e14676 Background: Ovarian cancer is the leading cause of death due to gynecologic malignancy. Biomarkers to predict chemoresponse and novel therapies to target these proteins would be practice changing. We aim to establish serum and tissue GAS6 as a predictive biomarker of chemoresponse and to determine if AXL inhibition through sequestration of its ligand, GAS6, with AVB-S6-500 (AVB) can improve chemoresponse. Methods: AVB was supplied by Aravive Biologics. High grade serous ovarian cancer (HGSOC) tumor samples were obtained pre- and post-neoadjuvant chemotherapy. AXL and GAS6 expression were evaluated by immunohistochemistry and serum concentration. In vitro viability and clonogenic assays were performed on chemoresistant tumor (OVCAR8, OVCAR5, COV62, and POV71-hTERT) and stromal cells (CAF86) treated with chemotherapy +/- AVB. Mouse models (OVCAR8, PDX, OVCAR5) were used to determine if the combination of chemotherapy + AVB reduced tumor burden. Immunofluorescent assays targeting ɣH2AX were used to evaluate DNA damage. Results: Patients with high pretreatment tumor GAS6 expression ( > 85%, n = 7) or serum GAS6 concentrations ( > 25ng/mL, n = 13) were more likely to be resistant to neoadjuvant chemotherapy than those with low tumor GAS6 expression ( < 45%, n = 4) (P = 0.010) or low serum GAS6 concentrations ( < 15ng/mL, n = 5) (P = 0.002). Carboplatin plus AVB (2µM, 5µM) and paclitaxel plus AVB (1µM) resulted in decreased cell viability and clonogenic growth compared to chemotherapy alone (p < 0.05) in all tumor and stromal cell lines. Synergism was seen between carboplatin+AVB and paclitaxel+AVB with a weighted combination index < 1. In vivo tumor mouse models treated with chemotherapy+AVB had significantly smaller subcutaneous and intraperitoneal (IP) tumors than those treated with chemotherapy alone (3.1mg vs 64mg, P = 0.003 OVCAR8; 62mg vs 157mg, P = 0.0108 PDX subcutaneous model; 0.05mg vs 0.3669mg, P < 0.001 OVCAR5 IP model). Increased DNA damage was noted in tumor and stromal cells treated with carboplatin+AVB compared to carboplatin alone (OVCAR8, COV362, CAF86 P < 0.001). Conclusions: High GAS6 is associated with lack of neoadjuvant chemoresponse in HGSOC patients. The combination of chemotherapy with AVB decreases tumor cell viability, tumor growth, and an increase in DNA damage response.

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Phage display screening identifies a novel peptide to suppress ovarian cancer cells in vitro and in vivo in mouse models

BMC Cancer ◽

10.1186/s12885-015-1891-8 ◽

2015 ◽

Vol 15 (1) ◽

Cited By ~ 7

Author(s):

Cong Zhou ◽

Jiali Kang ◽

Xiaoxia Wang ◽

Wei Wei ◽

Wenyan Jiang

Keyword(s):

Ovarian Cancer ◽

Phage Display ◽

Cancer Cells ◽

Mouse Models ◽

Ovarian Cancer Cells

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Quantification of Ultrasonic Scattering Properties of In Vivo Tumor Cell Death in Mouse Models of Breast Cancer

Translational Oncology ◽

10.1016/j.tranon.2015.11.001 ◽

2015 ◽

Vol 8 (6) ◽

pp. 463-473 ◽

Cited By ~ 16

Author(s):

Hadi Tadayyon ◽

Lakshmanan Sannachi ◽

Ali Sadeghi-Naini ◽

Azza Al-Mahrouki ◽

William T. Tran ◽

...

Keyword(s):

Breast Cancer ◽

Cell Death ◽

Tumor Cell ◽

Mouse Models ◽

Tumor Cell Death ◽

Ultrasonic Scattering

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Biomimetic Nanoparticles Blocking Autophagy for Enhanced Chemotherapy and Metastasis Inhibition via Reversing Focal Adhesion Disassembly

10.21203/rs.3.rs-917398/v1 ◽

2021 ◽

Author(s):

Yesi Shi ◽

Gan Lin ◽

Huili Zheng ◽

Dan Mu ◽

Hu Chen ◽

...

Keyword(s):

Tumor Cell ◽

Mouse Models ◽

Focal Adhesions ◽

Migration And Invasion ◽

Autophagic Flux ◽

Tumor Cell Migration ◽

Tumor Cell Motility ◽

Metastasis Inhibition

Abstract BackgroundAutophagy is a conserved catabolic process, which plays an important role in regulating tumor cell motility and degrading protein aggregates. Chemotherapy-induced autophagy may lead to tumor distant metastasis and even chemo-insensitivity in the therapy of hepatocellular carcinoma (HCC). However, a vast majority of HCC cases do not produce a significant response to monotherapy with autophagy inhibitors. ResultsIn this work, we develop a biomimetic co-delivery nanoformulation (TH-NP) co-encapsulating Oxaliplatin (OXA)/HCQ (hydroxychloroquine, an autophagy inhibitor) to execute targeted autophagy inhibition, reduce tumor cell migration and invasion in vitro and attenuate metastasis in vivo. Especially, TH-NPs can significantly improve OXA and HCQ concentration with approximately 21 and 13-fold increment in tumor tissues compared to the free mixture of HCQ/OXA. Moreover, the tumor-targeting TH-NPs release HCQ can alkalize the acidic lysosomes and thus inhibit the fusion of autophagosomes and lysosomes, leading to most effective blockade of autophagic flux compared to various controls. This largely improves chemotherapeutic performance of OXA in subcutaneous and orthotopic HCC mouse models. Importantly, TH-NPs also exhibit the most effective inhibition of tumor metastasis in orthotopic HCCLM3 models, and in the HepG2, Huh-7 or HCCLM3 metastatic mouse models. Then, we illustrate the enhanced metastasis inhibition is attributed to the blockade or reverse of the autophagy-mediated degradation of focal adhesions (FAs) including E-cadherin and paxillin. ConclusionsTH-NPs can perform an enhanced chemotherapy and antimetastatic effect, and may represent a promising strategy for HCC therapy in clinics.

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Tumorigenicity of Ascites-Derived Tumor Cells: Insights Into the Molecular Mechanisms of Ovarian Cancer Progression and Therapy Resistance

10.21203/rs.3.rs-117698/v1 ◽

2020 ◽

Author(s):

Yi Ding ◽

Vera Labitzky ◽

Karen Legler ◽

Minyue Qi ◽

Udo Schumacher ◽

...

Keyword(s):

Ovarian Cancer ◽

Tumor Cell ◽

Tumor Cells ◽

Cell Population ◽

Adherent Cell ◽

Cell Aggregates ◽

Tumorigenic Potential ◽

Extracellular Matrix Components

Abstract Background: Ovarian cancer (OvCa) cells disseminate primarily intraperitoneally. Here, detached tumor cell aggregates (spheroids) from the primary tumor are generally regarded as “metastatic units”, which exhibit a survival benefit, probably due to the protective microenvironment and their unique molecular characteristics. Hence, current therapeutic concepts such as classical chemotherapy are not sufficient preventing growth and spread of OvCa spheroids. Methods: In the current study we analyzed the cellular composition of ascites from ovarian cancer patients using flow cytometry and the tumorigenic potential of the different subpopulations in an intraperitoneal mouse model. Comparative transcriptome analyses (RNAseq) from ascites-derived tumor cells spheroids (n=10) vs. tumor samples from different metastatic sites (n=30) were further performed in order to identify key molecular players responsible for the special cellular characteristics of OvCa spheroids. Results: In vitro culture of ascites-derived cells gave rise to two different subpopulations: an adherent cell population (ADs) including mainly CD90+ cells with highly proliferative rates in vitro but no tumorigenic potential in vivo, and a non-adherent cell population (NADs) containing principally EpCAM+/CD24+ cells with low proliferative potential in vitro. NADs included cell aggregates and single cells, the first showing a high content (> 80%) of tumor cells (EpCAM+/CD24+). Enriched tumor cell spheroids from the ascites using cell strainers showed higher tumorigenic potential in vivo in comparison to the original ascites-derived cell population. Interestingly, the different metastatic spread patterns observed in the mice resembled the tumor dissemination pattern found in the corresponding patients. RNAseq analyses from tumor-spheroids revealed up-regulation of genes involved in chemoresistance (TGM1, HSPAs, MT1s), cell-adhesion and cell barrier (PKP3, CLDNs, PPL) and the oxidative phosphorylation (OXPHOS) process compared to the solid tumor tissue samples. Additionally, down-regulation of extracellular matrix components and angiogenesis-related genes could be observed. Targeting OXPHOS by metformin treatment led to reduced viability of ascites-derived spheroids from OvCa patients, showing to some extent a synergistic effect with cisplatin treatment. Conclusions: the actual study contributes to a better understanding of the biology of ovarian cancer spheroids and to the identification of new treatment opportunities in advanced ovarian cancer.

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Role of CD10 expression in endometriosis-associated mesenchymal stem cells on the progression of endometriosis-associated carcinoma.

Journal of Clinical Oncology ◽

10.1200/jco.2021.39.15_suppl.5561 ◽

2021 ◽

Vol 39 (15_suppl) ◽

pp. 5561-5561

Author(s):

Huda Atiya ◽

Taylor Orellana ◽

Lan Gardner Coffman

Keyword(s):

Ovarian Cancer ◽

Stem Cells ◽

Mesenchymal Stem Cells ◽

Cell Growth ◽

Tumor Cell ◽

Chemotherapy Resistance ◽

Cd10 Expression ◽

Sphere Formation

5561 Background: Endometriosis-associated carcinomas (EACs) such as ovarian clear cell cancer (OCCC) are rare, aggressive, chemo-resistant malignancies. While endometriosis is a known chronic inflammatory condition, the molecular mechanisms for the malignant transformation of endometriosis is unknown. Mesenchymal stem cells (MSC) are a critical component of the ovarian cancer microenvironment. Cancer cells reprogram MSCs to form carcinoma-associated MSCs (CAMSCs), which promote cancer growth, chemotherapy resistance, and metastases. MSCs are also found within the endometriotic microenvironment. CD10, a surface protein expressed by endometrial stromal cells, is also expressed on endometriosis-associated MSCs (enMSCs). Preliminary data demonstrate CD10 expression is lost in a subset of enMSCs and this loss is correlated with the acquisition of tumor-promoting properties. We hypothesized that the CD10 negative subset of enMSCs behave similarly to CAMSCs and support the growth of OCCC. Methods: EnMSCs were isolated from primary human benign endometriosis deposits involving the ovary or fallopian tubes. Flow cytometry was used to measure surface CD10 expression. We investigated the role of low CD10 enMSCs versus high CD10 enMSCs on OCCC tumor cell growth, chemotherapy resistance and stem-like cell properties in vitro and tumor cell engraftment, growth, and metastases in vivo. Luciferase-expressing OCCC cells were (1) used alone, (2) mixed with low CD10 enMSCs, or (3) mixed with high CD10 enMSCs and injected orthotopically into the ovarian bursa of NSG mice. In vivo imaging system was used to follow tumor progression and metastasis. Results: Our results demonstrated that enMSCs have variable CD10 expression. EnMSCs with low CD10 expression significantly enhanced OCCC proliferation, resistance to cisplatin, and sphere formation compared to OCCC alone. In contrast, high CD10 expressing enMSCs significantly reduce OCCC proliferation and sphere formation. Interestingly, low CD10 enMSCs selectively enhanced OCCC cell growth and had no effect on high grade serious ovarian cancer cell growth. Moreover, a reduction of CD10 expression was observed over time when high CD10 enMSCs were co-cultured with OCCC cells. Our results also showed enhanced tumor engraftment when OCCC cells were co-injected with low CD10 enMSCs to 100% one week post-injection, compared to 40% with OCCC and high CD10 enMSCs and 60% with OCCC alone. Further, mice co-injected with low CD10 enMSCs demonstrated increased metastasis and decreased survival compared to mice co-injected with high CD10 enMSCs. Conclusions: Our results indicate there is a sub population of enMSCs, marked by decreased CD10 expression, which selectively enhances OCCC growth. This highlights the existence of a tumor-promoting stromal cell within endometriosis which may be critical to the formation and propagation of EACs.

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PVT1, a YAP1 dependent stress responsive lncRNA drives ovarian cancer metastasis and chemoresistance

10.1101/2022.01.11.475893 ◽

2022 ◽

Author(s):

Kevin Tabury ◽

Mehri Monavarian ◽

Eduardo Listik ◽

Abigail K Shelton ◽

Alex Seok Choi ◽

...

Keyword(s):

Ovarian Cancer ◽

Tumor Cell ◽

Cell Survival ◽

Cancer Metastasis ◽

Cellular Stress ◽

Stress Factors ◽

Cellular Stress Response ◽

Ovarian Cancer Cells ◽

Dependent Manner

Metastatic growth of ovarian cancer cells into the peritoneal cavity requires adaptation to various cellular stress factors to facilitate cell survival and growth. Here we demonstrate the role of PVT1, one such stress induced long non-coding RNA, in ovarian cancer growth and metastasis. PVT1 is an amplified and overexpressed lncRNA in ovarian cancer with strong predictive value for survival and response to targeted therapeutics. We find that expression of PVT1 is regulated by ovarian tumor cells in response to cellular stress, particularly loss of cell-cell contacts and changes in matrix rigidity occurring in a YAP1 dependent manner. Induction of PVT1 promotes tumor cell survival, growth, and migration. Conversely, reducing PVT1 levels robustly abrogates metastatic behavior and tumor cell dissemination in cell lines and syngeneic transplantation models in vivo. We find that reducing PVT1 causes widespread transcriptome changes leading to alterations in cellular stress response and metabolic pathways including doxorubicin metabolism, which directly impacts chemosensitivity. Together, these findings implicate PVT1 as a promising therapeutic target to suppress metastasis and avoid chemoresistance in ovarian cancer.

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Gravity-based microfiltration reveals unexpected prevalence of circulating tumor cell clusters in ovarian cancer

10.1101/773507 ◽

2019 ◽

Author(s):

Anne Meunier ◽

Javier Alejandro Hernández-Castro ◽

Sara Kheireddine ◽

Sara Al Habyan ◽

Benjamin Péant ◽

...

Keyword(s):

Ovarian Cancer ◽

Tumor Cell ◽

Epithelial Ovarian Cancer ◽

Single Cell ◽

Tumor Cells ◽

Mouse Models ◽

Metastatic Potential ◽

Circulating Tumor Cell ◽

Large Size ◽

Localized Disease

SummaryCirculating tumor cells (CTCs) are rare (few cells per milliliter of blood) and mostly isolated as single cell CTCs (scCTCs). CTC clusters (cCTCs), even rarer, are of growing interest, notably because of their higher metastatic potential, but very difficult to isolate. Here, we introduce gravity-based microfiltration (GµF) for facile isolation of cCTCs. We identify cluster break-up as a confounding cause, and achieve ∼85% capture efficiency. GµF from orthotopic ovarian cancer mouse models and from 10 epithelial ovarian cancer (EOC) patients uncovered cCTCs in every case, with between 2-100+ cells. cCTCs represented between 5-30% of all CTC events, and 10-80% of captured CTCs were clustered; remarkably, in two patients, more CTCs were circulating as cCTCs than scCTCs. GµF uncovered the unexpected prevalence, frequency and sometimes large size of cCTCs in EOC patients with either metastatic and localized disease, and motivates additional studies to uncover their properties and role in disease progression.

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Modeling High-grade serous ovarian carcinoma using a combination of in vivo electroporation and CRISPR/Cas9 mediated genome editing

10.1101/2021.03.29.437221 ◽

2021 ◽

Author(s):

Katie Teng ◽

Matthew J Ford ◽

Keerthana Harwalkar ◽

YuQi Li ◽

David Farnell ◽

...

Keyword(s):

Ovarian Cancer ◽

Ovarian Carcinoma ◽

Genome Editing ◽

Mouse Models ◽

Gynecological Cancer ◽

Lineage Tracing ◽

High Grade ◽

In Vivo Electroporation ◽

Serous Ovarian Carcinoma

Ovarian cancer remains the most lethal gynecological cancer today. High-grade serous ovarian carcinoma (HGSC) is the most common and lethal type of ovarian cancer and is most frequently diagnosed at advanced stages. Here, we developed a novel strategy to generate somatic ovarian cancer mouse models using a combination of in vivo electroporation and CRISPR/Cas9 mediated genome editing. We mutated tumor suppressor genes associated with HGSC in two different combinations; Brca1, Tp53, Pten with/without Lkb1 and successfully generated HGSC, however, with different latencies and pathophysiology. By utilizing Cre lineage tracing in our system, we visualized peritoneal micrometastases in an immune-competent environment. Because our strategy is flexible in selecting mutation combinations and targeting areas, it would be useful for generating ovarian cancer mouse models.

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