Micron-sized and submicron-sized aerosol deposition in a new ex vivo preclinical model

AbstractBreast cancer is now globally the most frequent cancer and leading cause of women’s death. Two thirds of breast cancers express the luminal estrogen receptor-positive (ERα + ) phenotype that is initially responsive to antihormonal therapies, but drug resistance emerges. A major barrier to the understanding of the ERα-pathway biology and therapeutic discoveries is the restricted repertoire of luminal ERα + breast cancer models. The ERα + phenotype is not stable in cultured cells for reasons not fully understood. We examine 400 patient-derived breast epithelial and breast cancer explant cultures (PDECs) grown in various three-dimensional matrix scaffolds, finding that ERα is primarily regulated by the matrix stiffness. Matrix stiffness upregulates the ERα signaling via stress-mediated p38 activation and H3K27me3-mediated epigenetic regulation. The finding that the matrix stiffness is a central cue to the ERα phenotype reveals a mechanobiological component in breast tissue hormonal signaling and enables the development of novel therapeutic interventions. Subject terms: ER-positive (ER + ), breast cancer, ex vivo model, preclinical model, PDEC, stiffness, p38 SAPK.

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Evaluation of ON 01910.Na, a novel modulator of polo-like kinase 1 (Plk1) pathway, and development of a cyclin-B1-based predictive assay in pancreatic cancer

Journal of Clinical Oncology ◽

10.1200/jco.2007.25.18_suppl.3569 ◽

2007 ◽

Vol 25 (18_suppl) ◽

pp. 3569-3569

Author(s):

A. Jimeno ◽

A. Chan ◽

X. Zhang ◽

J. Wheelhouse ◽

A. Solomon ◽

...

Keyword(s):

Pancreatic Cancer ◽

Cell Lines ◽

Ex Vivo ◽

Cyclin B1 ◽

Preclinical Model ◽

Significant Activity ◽

In Vivo Models ◽

Pancreatic Cell ◽

Ex Vivo Assay

3569 Background: Plk1 is a key mitotic regulator of the transition through the G2/M checkpoint in the cell cycle. This work aimed to evaluate the activity of ON 01910.Na, a Plk1 pathway modulator, in in vitro and in vivo models of pancreatic cancer (PaCa) and to discover biomarkers predictive of efficacy. Methods: ON 01910.Na was tested in 12 PaCa cell lines. Studies assessing Plk1 related markers were conducted to identify biomarkers. For validation a live collection of PaCa xenografts from fresh tumor samples obtained at the time of surgical resection was used (PancXenoBank). The ex vivo assay was based on fine-needle aspirate (FNA) biopsies. Results: ON 01910.Na showed equal activity to gemcitabine against PaCa cell lines. The activity of ON 01910.Na correlated with suppression of two downstream mediators of PLK1, CDC25C and cyclin B1 (by mRNA and protein). ON 01910.Na was tested in xenografts from representative pancreatic cell lines. The selected markers were evaluated in an ex vivo assay, using intra-tumor pharmacokinetics to select the dose of the assay. Cyclin B1 mRNA evaluation yielded the most optimal combination of accuracy and reproducibility. Knockdown of cyclin B1 by siRNA had no effect per se or in the response of the resistant MiaPaca2 to either of the drugs. We next used the ex vivo assay to profile ten patient-derived cases from the PancXenoBank. Two cases were catalogued as potential responders. From each of these ten cases, a group of mice bearing at least 20 tumors received vehicle or ON 01910.Na for 28 days. There was a correlation between the ex vivo cyclin B1 assay and the sensitivity to the tested agent, as the 2 cases prospectively identified as sensitive met pre-specified criteria for response. Of the 8 tumors predicted to be resistant, only one was sensitive. In IHC testing cases showing ex vivo cyclin B1 down-regulation had also decreases in cyclin B1 protein, and there was a correlation between activity and IHC changes in cyclin B1. Conclusions: ON 01910.Na demonstrated significant activity in a preclinical model of PaCa. A rationally designed ex vivo cyclin B1-based assay not only identified cases sensitive to ON 01910.Na, but also replicated the pharmacodynamic events occurring after in vivo exposure. No significant financial relationships to disclose.

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A unique three-dimensional model for evaluating the impact of therapy on multiple myeloma

Blood ◽

10.1182/blood-2008-02-142430 ◽

2008 ◽

Vol 112 (7) ◽

pp. 2935-2945 ◽

Cited By ~ 73

Author(s):

Julia Kirshner ◽

Kyle J. Thulien ◽

Lorri D. Martin ◽

Carina Debes Marun ◽

Tony Reiman ◽

...

Keyword(s):

Stem Cells ◽

Multiple Myeloma ◽

Plasma Cells ◽

Ex Vivo ◽

Three Dimensional ◽

Preclinical Model ◽

Drug Resistant ◽

Three Dimensional Model ◽

The Impact

AbstractAlthough the in vitro expansion of the multiple myeloma (MM) clone has been unsuccessful, in a novel three-dimensional (3-D) culture model of reconstructed bone marrow (BM, n = 48) and mobilized blood autografts (n = 14) presented here, the entire MM clone proliferates and undergoes up to 17-fold expansion of malignant cells harboring the clonotypic IgH VDJ and characteristic chromosomal rearrangements. In this system, MM clone expands in a reconstructed microenvironment that is ideally suited for testing specificity of anti-MM therapeutics. In the 3-D model, melphalan and bortezomib had distinct targets, with melphalan targeting the hematopoietic, but not stromal com-partment. Bortezomib targeted only CD138+CD56+ MM plasma cells. The localization of nonproliferating cells to the reconstructed endosteum, in contact with N-cadherin–positive stroma, suggested the presence of MM-cancer stem cells. These drug-resistant CD20+ cells were enriched more than 10-fold by melphalan treatment, exhibited self-renewal, and generated clonotypic B and plasma cell progeny in colony forming unit assays. This is the first molecularly verified demonstration of proliferation in vitro by ex vivo MM cells. The 3-D culture provides a novel biologically relevant preclinical model for evaluating therapeutic vulnerabilities of all compartments of the MM clone, including presumptive drug-resistant MM stem cells.

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Aerosol delivery during invasive mechanical ventilation: development of a preclinical ex vivo respiratory model for aerosol regional deposition

Scientific Reports ◽

10.1038/s41598-019-54480-9 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Yoann Montigaud ◽

Quentin Georges ◽

Jérémie Pourchez ◽

Lara Leclerc ◽

Clémence Goy ◽

...

Keyword(s):

Mechanical Ventilation ◽

Ex Vivo ◽

Imaging Techniques ◽

Invasive Mechanical Ventilation ◽

Cost Effective ◽

Aerosol Deposition ◽

Regional Deposition ◽

Porcine Respiratory ◽

Good Agreement

AbstractIn intensive care units, nebulization is a usual route for drug administration to patients under mechanical ventilation (MV). The effectiveness of inhalation devices as well as depositions sites of aerosols for ventilated patients remain poorly documented. In vivo human inhalation studies are scarce due to ethical restrictions because imaging techniques require radioaerosols to assess regional aerosol deposition. Thus, we developed an ex vivo respiratory model under invasive MV for preclinical aerosol deposition studies. The model was composed of ex vivo porcine respiratory tracts. MV was achieved thanks to a tracheal intubation and a medical ventilator under controlled conditions. Respiratory features were studied using analogical sensors. Then regional homogeneity of gas-ventilation was assessed with 81mKrypton scintigraphies. Finally, a proof of concept study for aerosol deposition was performed. Obtained respiratory features as well as gamma-imaging techniques, which demonstrated a homogenous regional ventilation and about 18% ± 4% of the nebulized dose deposited the respiratory tract, were in good agreement with human data available in the literature. This original ex vivo respiratory model provides a feasible, reproducible and cost-effective preclinical tool to achieve aerosol deposition studies under MV.

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P2.03-17 Optimization of an Ex-Vivo Preclinical Model for Drug Testing

Journal of Thoracic Oncology ◽

10.1016/j.jtho.2019.08.1464 ◽

2019 ◽

Vol 14 (10) ◽

pp. S689

Author(s):

E. Marin ◽

P. Duch ◽

M. Gabasa ◽

E. Urrea ◽

R. Ikemori ◽

...

Keyword(s):

Drug Testing ◽

Ex Vivo ◽

Preclinical Model

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Organoids in Lung Cancer Management

Frontiers in Surgery ◽

10.3389/fsurg.2021.753801 ◽

2021 ◽

Vol 8 ◽

Author(s):

Yushi Li ◽

Joyce W. Y. Chan ◽

Rainbow W. H. Lau ◽

Winnie W. Y. Cheung ◽

Alissa Michelle Wong ◽

...

Keyword(s):

Lung Cancer ◽

Lung Tumor ◽

Ex Vivo ◽

Three Dimensional ◽

Dimensional Structure ◽

Patient Specific ◽

Preclinical Model ◽

Cancer Tissue ◽

Cancer Management

Lung cancer is a complex milieu of genomically altered cancer cells, a diverse collection of differentiated cells and nonneoplastic stroma. Lung cancer organoids is a three-dimensional structure grown from patient cancer tissue that could mimic in vivo complex behavior and cellular architecture of the cancer. Furthermore, the genomic alterations of the primary lung tumor is captured ex vivo. Lung cancer organoids have become an important preclinical model for oncology studies in recent years. It could be used to model the development of lung cancer, investigate the process of tumorigenesis, and also study the signaling pathways. The organoids could also be a platform to perform drug screening and biomarker validation of lung cancer, providing a promising prediction of patient-specific drug response. In this review, we described how lung cancer organoids have opened new avenues for translating basic cancer research into clinical therapy and discussed the latest and future developments in organoid technology, which could be further applied in lung cancer organoids research.

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Ex-Vivo Treatment of Tumor Tissue Slices as a Predictive Preclinical Method to Evaluate Targeted Therapies for Patients with Renal Carcinoma

Cancers ◽

10.3390/cancers12010232 ◽

2020 ◽

Vol 12 (1) ◽

pp. 232 ◽

Cited By ~ 7

Author(s):

Caroline Roelants ◽

Catherine Pillet ◽

Quentin Franquet ◽

Clément Sarrazin ◽

Nicolas Peilleron ◽

...

Keyword(s):

Targeted Therapies ◽

Tumor Tissue ◽

Ex Vivo ◽

Human Tumor ◽

New Drugs ◽

High Rate ◽

Preclinical Model ◽

Tissue Slices ◽

Cell Renal Cell Carcinoma ◽

Correct Treatment

Clear cell renal cell carcinoma (ccRCC) is the third type of urologic cancer. At time of diagnosis, 30% of cases are metastatic with no effect of chemotherapy or radiotherapy. Current targeted therapies lead to a high rate of relapse and resistance after a short-term response. Thus, a major hurdle in the development and use of new treatments for ccRCC is the lack of good pre-clinical models that can accurately predict the efficacy of new drugs and allow the stratification of patients into the correct treatment regime. Here, we describe different 3D cultures models of ccRCC, emphasizing the feasibility and the advantage of ex-vivo treatment of fresh, surgically resected human tumor slice cultures of ccRCC as a robust preclinical model for identifying patient response to specific therapeutics. Moreover, this model based on precision-cut tissue slices enables histopathology measurements as tumor architecture is retained, including the spatial relationship between the tumor and tumor-infiltrating lymphocytes and the stromal components. Our data suggest that acute treatment of tumor tissue slices could represent a benchmark of further exploration as a companion diagnostic tool in ccRCC treatment and a model to develop new therapeutic drugs.

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Thrombogenicity of flow diverters in an ex vivo shunt model: effect of phosphorylcholine surface modification

Journal of NeuroInterventional Surgery ◽

10.1136/neurintsurg-2016-012612 ◽

2016 ◽

Vol 9 (10) ◽

pp. 1006-1011 ◽

Cited By ~ 34

Author(s):

Matthew W Hagen ◽

Gaurav Girdhar ◽

John Wainwright ◽

Monica T Hinds

Keyword(s):

Surface Modification ◽

Antiplatelet Therapy ◽

Ex Vivo ◽

Cerebral Aneurysms ◽

Flow Diverter ◽

Preclinical Model ◽

Platelet Deposition ◽

Treatment For Cerebral Aneurysms ◽

Flow Diverters

BackgroundFlow diverters offer a promising treatment for cerebral aneurysms. However, they have associated thromboembolic risks, mandating chronic dual antiplatelet therapy (DAPT). Shield Technology is a phosphorylcholine surface modification of the Pipeline Embolization Device (PED) flow diverter, which has shown significant reductions in material thrombogenicity in vitro.ObjectiveTo compare the thrombogenicity of PED, PED with Shield Technology (PED+Shield), and the Flow-Redirection Endoluminal Device (FRED)—with and without single antiplatelet therapy and DAPT—under physiological flow.MethodsAn established non-human primate ex vivo arteriovenous shunt model of stent thrombosis was used. PED, PED+Shield, and FRED were tested without antiplatelet therapy, with acetylsalicylic acid (ASA) monotherapy, and with DAPT. Radiolabeled platelet deposition was quantified over 1 hour for each device and total fibrin deposition was also quantified.ResultsCumulative statistical analysis showed significantly lower platelet deposition on PED compared with FRED. The same statistical model showed significant decreases in platelet deposition when ASA, clopidogrel, or Shield Technology was used. Direct comparisons of device performances within antiplatelet conditions showed consistent significant decreases in platelet accumulation on PED+Shield relative to FRED. PED+Shield showed significant reductions in platelet deposition compared with unmodified PED without antiplatelet therapy and with DAPT. PED accumulated minimal fibrin with and without Shield Technology.ConclusionsIn this preclinical model, we have shown that the Shield Technology phosphorylcholine modification reduces the platelet-specific thrombogenicity of a flow diverter under physiologically relevant flow with and without DAPT. We have further identified increased fibrin-driven thrombogenicity associated with FRED relative to PED.

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