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.

scholarly journals A microfluidic platform for cultivating ovarian cancer spheroids and testing their responses to chemotherapies

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Neda Dadgar ◽  
Alan M. Gonzalez-Suarez ◽  
Pouria Fattahi ◽  
Xiaonan Hou ◽  
John S. Weroha ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Cancer Cells ◽  
Drug Testing ◽  
Microfluidic Devices ◽  
Needle Aspiration ◽  
High Concentration ◽  
Microfluidic Platform ◽  
Drug Concentrations

Abstract There is increasing interest in utilizing in vitro cultures as patient avatars to develop personalized treatment for cancer. Typical cultures utilize Matrigel-coated plates and media to promote the proliferation of cancer cells as spheroids or tumor explants. However, standard culture conditions operate in large volumes and require a high concentration of cancer cells to initiate this process. Other limitations include variability in the ability to successfully establish a stable line and inconsistency in the dimensions of these microcancers for in vivo drug response measurements. This paper explored the utility of microfluidics in the cultivation of cancer cell spheroids. Six patient-derived xenograft (PDX) tumors of high-grade serous ovarian cancer were used as the source material to demonstrate that viability and epithelial marker expression in the microfluidic cultures was superior to that of Matrigel or large volume 3D cultures. To further demonstrate the potential for miniaturization and multiplexing, we fabricated multichamber microfluidic devices with integrated microvalves to enable serial seeding of several chambers followed by parallel testing of several drug concentrations. These valve-enabled microfluidic devices permitted the formation of spheroids and testing of seven drug concentrations with as few as 100,000 cancer cells per device. Overall, we demonstrate the feasibility of maintaining difficul-to-culture primary cancer cells and testing drugs in a microfluidic device. This microfluidic platform may be ideal for drug testing and personalized therapy when tumor material is limited, such as following the acquisition of biopsy specimens obtained by fine-needle aspiration.

scholarly journals XenoSarc: A comprehensive platform of patient-derived xenograft (PDX) models of soft tissue sarcoma (STS) for early drug testing.

2019 ◽  
Vol 5 (suppl) ◽  
pp. 37-37 ◽  
Author(s):  
Patrick Schoffski ◽  
Britt Van Renterghem ◽  
Jasmien Cornillie ◽  
Yannick Wang ◽  
Yemarshet Kelemework Gebreyohannes ◽  
...  
Keyword(s):  
Drug Testing ◽  
Treatment Options ◽  
Clinical Information ◽  
Tissue Microarrays ◽  
Mouse Tissue ◽  
Tissue Samples ◽  
Pdx Models ◽  
Early Drug

37 Background: STS is a family of rare, heterogeneous tumors with > 70 subtypes. There is an urgent need for reliable preclinical models, especially for orphan subtypes of STS, given the limited treatment options. Methods: A panel of PDX models was established by s.c. implantation of fresh tumor specimens in athymic NMRI mice. Growing pieces of tumor were re-transplanted to next generations of mice. At each passage fragments were collected for histological/molecular characterization. A model was considered “established” after observing stable features for at least 2 passages. Ex-mouse tissue samples were stored, characterized by immunohistochemistry/flow cytometry and used for in vitro drug testing. Results: Between 2011-2019, 329 samples from 301 consenting patients were transplanted; 56 models are established, 16 additional models are in early passaging. Clinical information about donor and tumor (including sensitivity to standard and experimental agents) is available. The platform includes models of dedifferentiated lipo- (10 models), myxofibro- (8), leiomyo- (7), synovial (2), intimal (2), CIC-positive round cell (1), mesenchymal chondro- (1), extraskeletal osteo- (1), myxoid lipo- (1), myxoinflammatory fibroblastic (1), rhabdomyo- (2) and high-grade undifferentiated pleomorphic sarcoma (7), as well as GIST (8), MPNST (4) and epithelioid hemangioendothelioma (1). Models are well-characterized, with molecular information on copy number changes (low-coverage whole genome sequencing) and gene expression profile (RNA-Seq) available. We also constructed tissue microarrays from the xenografts which are used for target identification and model selection for preclinical studies. Xenografts are available for in vivo testing of novel agents, and results already served as a rationale for a number of prospective clinical trials. Conclusions: XenoSarc offers opportunities for studying the biology of a variety of sarcoma subtypes including ultra-rare entities and is a valuable tool for early drug screening in preparation of clinical STS trials. The platform is well maintained and continuously expanded, and available to collaborators from academia and industry.

scholarly journals Results of CoALL 07-03 study childhood ALL based on combined risk assessment by in vivo and in vitro pharmacosensitivity

2019 ◽  
Vol 3 (22) ◽  
pp. 3688-3699 ◽  
Author(s):  
Franziska Schramm ◽  
Udo zur Stadt ◽  
Martin Zimmermann ◽  
Norbert Jorch ◽  
Arnulf Pekrun ◽  
...  
Keyword(s):  
Drug Testing ◽  
Drug Response ◽  
Lymphoblastic Leukemia ◽  
Term Outcome ◽  
Childhood All ◽  
Long Term Outcome ◽  
Key Points

Key Points Report of the long-term outcome of children with acute lymphoblastic leukemia upon risk-adapted therapy accrued in trial CoALL 07-03. Lack of correlation between in vitro and in vivo drug response as well as a lower predictive value of in vitro drug testing.

scholarly journals Assessment of an in vitro physiological relevant model to check therapeutic strategies for glaucoma

2020 ◽  
Author(s):  
Sara Tirendi ◽  
Stefania Vernazza ◽  
Sergio Saccà ◽  
Anna Maria Bassi
Keyword(s):  
Drug Testing ◽  
Peripheral Vision ◽  
Early Stage ◽  
Treatment And Prevention ◽  
Chronic Oxidative Stress ◽  
Repeatability And Reproducibility ◽  
Vitro Model ◽  
Key Events

Glaucoma is a chronic, progressive and heterogeneous optic neuropathy which affects in the early stage the peripheral vision and then the central vision, leading to irreversible blindness. As known, in glaucoma the trabecular meshwork represent the main tissue which is impaired by chronic oxidative stress, aging and increase of intraocular pressure. Today, the lack of human-based models, with characteristics of high repeatability and reproducibility as well, called for an high-quality in vitro model with a good degree of resemblance for the tissue or organ of interest as a basis for new drug testing. Our team has been committed to this purpose by assessment of an in vitro 3D TM human-based model, closer to in vivo, using millifluidic technology, to better identify the key events underlying the pathogenesis of glaucoma and to evaluate new therapies targeted at disease treatment and prevention.

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.

Opening New Horizons in the Treatment of Childhood Onset Leukodystrophies

2019 ◽  
Vol 50 (04) ◽  
pp. 211-218 ◽  
Author(s):  
Stina Schiller ◽  
Marco Henneke ◽  
Jutta Gärtner
Keyword(s):  
Early Stage ◽  
Treatment Options ◽  
Clinical Signs ◽  
Disease Genes ◽  
Lysosomal Storage ◽  
Peroxisomal Disorders ◽  
Enzyme Replacement ◽  
Number Of Patients

AbstractLeukodystrophies (LDs) predominantly affect the white matter of the central nervous system and its main component, the myelin. The majority of LDs manifests in infancy with progressive neurodegeneration. Main clinical signs are intellectual and motor function losses of already attained developmental skills. Classical LDs include lysosomal storage disorders like metachromatic leukodystrophy (MLD), peroxisomal disorders like X-linked adrenoleukodystrophy (X-ALD), disorders of mitochondrial dysfunction, and myelin protein defects like Pelizaeus-Merzbacher disease. So far, there are only single LD disorders with effective treatment options in an early stage of disease. The increasing number of patients diagnosed with LDs emphasizes the need for novel therapeutic options. Impressive advances in biotechnology have not only led to the continuous identification of new disease genes for so far unknown LDs but also led to new effective neuroprotective and disease-modifying therapeutic approaches. This review summarizes ongoing and novel innovative treatment options for LD patients and their challenges. It includes in vitro and in vivo approaches with focus on stem cell and gene therapies, intrathecal substrate or enzyme replacement, and genome editing.

scholarly journals ETMR-18. TARGETING Lin28 IN ETMR WITH ODC1 INHIBITOR DFMO

2020 ◽  
Vol 22 (Supplement_3) ◽  
pp. iii326-iii326
Author(s):  
Austin Goodyke ◽  
Shannon Kelly ◽  
Joseph Zagorski ◽  
Elizabeth VanSickle ◽  
Tyler Maser ◽  
...  
Keyword(s):  
Drug Testing ◽  
Treatment Options ◽  
Young Patients ◽  
Treatment Strategies ◽  
Therapeutic Agents ◽  
Tumor Burden ◽  
Protein Levels ◽  
New Treatment

Abstract Embryonal tumor with multilayered rosettes (ETMR), is an aggressive brain tumor primarily occurring in young patients (<4 years of age) and characterized by C19MC amplification and Lin28 overexpression. These genetic hallmarks have been shown to participate in driving ETMR in a C19MC-Lin28-MYCN circuit. Reducing Lin28 disrupts this circuit and reduces cell viability in ETMR models. Investigation of therapeutic agents targeting this pathway is required to provide new treatment options for this deadly disease. We present data showing the effect of DFMO (α-difluoromethylornithine) in ETMR, an ODC1 inhibitor known to reduce Lin28 in neuroblastoma. DFMO treatment of the ETMR cell line BT-183 resulted in a significant reduction of intracellular Lin28 protein levels (P<0.05) as indicated by flow cytometry. In concert with this reduction in Lin28, there was a as significant reduction in viable cells (P<0.05), and the number of CD133+ cells were reduced 2-fold (P<0.05). High throughput drug testing of BT-183 identified a number of additional therapeutic agents with potential therapeutic efficacy for ETMR and combining these with cytostatic agent DFMO demonstrated the potential use of these drugs in combination. These in vitro data were complemented by testing of DFMO in an in vivo stereotaxic xenograft ETMR model, with inhibition of tumor burden monitored by bioluminescent imaging of the tumors. Together this work shows that Lin28 targeting agents such as DFMO merit further examination and integrating these types of agents into treatment strategies for ETMR may lead to better outcomes.

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.

scholarly journals 47. UNCOVERING A NOVEL ROLE FOR HLA-G IN BRAIN METASTASES

2020 ◽  
Vol 2 (Supplement_2) ◽  
pp. ii9-ii9
Author(s):  
Blessing Bassey-Archibong ◽  
Nikoo Aghaei ◽  
Chirayu Chokshi ◽  
Agata Kieliszek ◽  
Nazanin Tatari ◽  
...  
Keyword(s):  
Brain Metastases ◽  
Early Stage ◽  
Treatment Options ◽  
Current Treatment ◽  
Primary Brain Tumours ◽  
Metastatic Cells ◽  
Mature Brain ◽  
Pdx Models

Abstract Brain metastases (BM) are the most common brain tumour in adults and are ten times more likely to develop than primary brain tumours. More than 20% of patients with cancer will develop BM with the three most common sources being primary cancers of the lung, breast, and melanoma. Unfortunately, current treatment options for BM do not effectively eradicate BM, with a mere median overall survival time of 12 months in treated patients. This indicates the need for better and more effective therapies against BM. Using patient-derived cell lines established from surgically removed brain metastatic tumours of lung-, breast- and melanoma-BM patients, we generated patient-derived orthotopic murine xenograft (PDX) models of lung-, breast-, and melanoma-BM. From these PDX models, we isolated a rare population of stem-like brain metastasis initiating cells (BMICs) we termed “pre-metastatic”, that had traveled from their primary/orthotopic tumours and lodged in the brain but had not yet developed into mature BM. Transcriptomic analyses performed on pre-metastatic and non-pre-metastatic BMICs from lung, breast and melanoma PDX models of BM, identified a set of deregulated genes exclusive only to pre-metastatic BMICs. Further analysis revealed HLA-G as being commonly up-regulated only during the pre-metastatic stage of the lung-, breast-, and melanoma-BM cascade. In vitro and in vivo analyses demonstrated that HLA-G knock-down reduced the proliferation and survival of BMICs from all BM cohorts, and attenuated the establishment of mature brain metastatic tumours, implying a crucial role for HLA-G in the formation of BM. Developing a therapeutic strategy that targets HLA-G in BM may prove effective at completely eliminating brain metastatic cells at an early stage of the BM cascade, thereby turning a fatal disease into an eminently more treatable one.

scholarly journals Mechanism of CK2.3, a Novel Mimetic Peptide of Bone Morphogenetic Protein Receptor Type IA, Mediated Osteogenesis

2019 ◽  
Vol 20 (10) ◽  
pp. 2500 ◽  
Author(s):  
Vrathasha Vrathasha ◽  
Hilary Weidner ◽  
Anja Nohe
Keyword(s):  
Signaling Pathways ◽  
Treatment Options ◽  
Time Frame ◽  
Bmp Signaling ◽  
C2c12 Cells ◽  
Molecular Sequence ◽  
Sequence Of Events ◽  
Protein Receptor

Background: Osteoporosis is a degenerative skeletal disease with a limited number of treatment options. CK2.3, a novel peptide, may be a potential therapeutic. It induces osteogenesis and bone formation in vitro and in vivo by acting downstream of BMPRIA through releasing CK2 from the receptor. However, the detailed signaling pathways, the time frame of signaling, and genes activated remain largely unknown. Methods: Using a newly developed fluorescent CK2.3 analog, specific inhibitors for the BMP signaling pathways, Western blot, and RT-qPCR, we determined the mechanism of CK2.3 in C2C12 cells. We then confirmed the results in primary BMSCs. Results: Using these methods, we showed that CK2.3 stimulation activated OSX, ALP, and OCN. CK2.3 stimulation induced time dependent release of CK2β from BMPRIA and concurrently CK2.3 colocalized with CK2α. Furthermore, CK2.3 induced BMP signaling depends on ERK1/2 and Smad1/5/8 signaling pathways. Conclusion: CK2.3 is a novel peptide that drives osteogenesis, and we detailed the molecular sequence of events that are triggered from the stimulation of CK2.3 until the induction of mineralization. This knowledge can be applied in the development of future therapeutics for osteoporosis.

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