Ovo Balance Enhancer Application – Evaluation of angiogenesis of polyhydroxybutyric acid and chitosan-based acellular porous biomaterials using the chicken extraovarian intestinal urinary membrane model.
A real-world multicenter collaborative retrospective study of low-dose afatinib for human epidermal growth factor receptor 2-negative metastatic breast cancer.
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Chicken embryonic tumor xenografts derived from circulating tumor cells as a relevant model to study the spread of metastases: a proof of concept.
Xavier Rousset, David Barthélemy David Barthélemy Skillit Preprints.org Google Scholar View Publications 7, 8, Sara Calatini Sara Calatini Skillit Preprints.org Google Scholar View Publications 9, Marie Brevet Marie Brevet Skillit Preprints.org Google Scholar View Publications Julie Balandier Skillit Preprints. org Google Scholar Views Publications 7, 8, Margaux Raffin Margaux Raffin Skillit Preprints.org Google Scholar Views Publications 7, 8, Florence Giguère Florence Giguère Skillit Preprints.org Google Scholar Views Publications 7, 8, J Skillit Preprints. Organization Google Scholar Views Publications 7, 8, Myriam Decoussin-Petruucci Myriam Decoussin-Petruucci Skillit Preprints.org Google Scholar Views Publications 2, 6, 11, Julien Perron Julien Perron Skillit Preprints.org Google Scholar Views Publicaz 2, 1 Benzardjeb Skillit Preprints. org Google Scholar View Publications 2, 6, 11, Sébastien Couraud Sébastien Couraud Skillit Preprints.org Google Scholar View Publications 2, 5, 6, Jean Viallet Jean Viallet Skillit Preprints.org Google Scholar View Publications 1 and Payne Skillit Preprints.org Google Scholar View Publications 2, 6, 7, 8, *
Laboratoire de Biométrie et Biologie Evolutive, Equipe Biostatistique-Santé, CNRS UMR 5558, Université Claude Bernard Lyon 1, 69100 Villeurbanne, France.
Receipt date: June 10, 2022 / Revision date: July 21, 2022 / Repair date: August 19, 2022 / Publication date: August 23, 2022
Phosphatidylcholine: Beyond Cellular Integrity
(This article belongs to the special issue Choriourea (CAM) Model – Traditional and State-of-the-Art Applications: 1st International CAM Conference)
Circulating tumor cells (CTCs) are heterogeneous in the bloodstream and, in rare cases, are a source of cancer metastasis. Such expansion in vitro or in vivo remains a great challenge. Chicken enterocyst (CAM) assay has proven to be a reliable alternative to mouse models, especially in tumor xenografts. We developed a promising model of CTC-derived xenografts in chicken CAM and demonstrated that next-generation sequencing (NGS) analysis is feasible in this assay along with genomic match between in-vitro tumors and the original patient’s tumor. We also demonstrated the spread of metastases from chicken embryo xenografts to distant organs. Further characterization of in vitro tumors and metastases may provide new insights into mechanisms of tumor dissemination. Developing xenografts from a specific patient’s CTCs within a time frame compatible with the patient’s therapeutic management could also be a step toward individualized treatment.
Patient-derived xenografts (PDXs) of enterocysts (CAMs) are representative models for human tumor research. Circulating tumor cells (CTC) are involved in cancer proliferation and treatment resistance mechanisms. Much effort has been made to expand the cells to facilitate the study and detailed analysis of these small numbers of cells. Here, we evaluated whether isolation of fresh CTCs from patients with metastatic cancer can provide a reliable tumor model after CAM xenografting. We enrolled 35 patients with breast cancer, prostate cancer, and lung metastasis cancer. Microfluidic-based CTC culture was performed. After 48–72 h of incubation, CTCs were seeded into the CAM of developing chicken eggs at day 9 of embryogenesis (EDD9). Nine days after transplantation, tumors were resected and histopathological, immunochemical, and genomic analyzes were performed. Intraovarian tumors were obtained in 61% of patients. Various types of small tumors with spindle-shaped cells were observed. The epithelial-to-mesenchymal transition of CTCs may explain this phenotype. Beyond the feasibility of NGS in this model, we highlighted the genomic concordance between the intraovarian tumor and the original patient’s tumor with regard to constitutional polymorphisms and somatic changes in one patient. Alu DNA sequences were detected in isolated organs of chicken embryos and confirmed the idea of differentiated cells with aggressive behavior. To the best of our knowledge, we have demonstrated the first chicken CAM CTC-derived xenograft using NGS analysis and evidence of CTC proliferation in chicken embryos.
Cancer remains the leading cause of death worldwide, particularly in the lung, prostate, and breast [1]. In an aging society, the burden of cancer incidence and mortality is rapidly increasing, and multiple exposures to various cancer risk factors are increasing. This has led to intensive scientific research and many advances in understanding the molecular mechanisms. Recently, there have been significant improvements in the diagnosis and management of this heterogeneous disease [2]. Despite these recent advances, turnover rates in drug development remain high. Therefore, extensive improvements in in vivo models are needed to provide optimal treatment options for patients and facilitate novel drug discovery. Patient tumor models in mice are one of the most widely used models. Although these models have many advantages, they are neither cost-effective nor time-efficient in the case of personalized medicine and cannot predict the efficacy of a treatment for a specific patient within the treatment window. [3].
Chx10 Consolidates V2a Interneuron Identity Through Two Distinct Gene Repression Modes
Driven by the increase in personalized medicine and the global demand for compliance with the Reduce, Exchange, and Purify (3R) policy, alternative models, such as chicken urinary membrane (CAM) xenografts, are being developed. The CAM is composed of the chorionic epithelium, mesodermal layer, and allantoic epithelium. This embryonic membrane is connected to the embryo through a continuous embryonic vasculature and is easily accessible for manipulation and observation [4]. The highly vascularized membrane and the natural immune dysfunction upon attachment (EDD9 phase) provide an environment rich in nutrients and embryonic growth factors, which favors aggressive tumor proliferation. The in ovo model is well described to mimic intratumoral hypoxia [5]. Xenografts from cell lines to patient tumor-derived xenografts have been performed for decades for many types of cancer in vivo [6, 7, 8]. Comparison of CAM and patient tumors showed good similarity in both histological and immunochemical analyses. Analysis of metastatic spread, angiogenesis, tumorigenesis and drug chemosensitivity tests have been shown to be reliable [9, 10, 11]. For example, in 1991, Shoen et al. Twenty-one xenografts from untreated malignant gliomas were developed into CAM. The xenograft and patient were treated with anticancer drugs. There was 78% agreement between in ovo responses and corresponding patient responses [12]. Rapid readout reduces the gap between assessment and assessment of therapeutic effectiveness on xenografts and greatly improves drug development.
Circulating cancer biomarkers, such as circulating tumor DNA (ctDNA) and circulating tumor cells (CTC), are emerging as valuable resources for non-invasive diagnosis, prognostic assessment, treatment monitoring, and outcome prediction [13]. Importantly, 25% to 67% of patients with non-small cell lung cancer (NSCLC) do not benefit from repeat biopsy during early progression. Even if regenerative testing is possible, molecular interpretation of the results is limited by the spatial and temporal heterogeneity of the tumor [14]. CTC exploration provides a snapshot of tumor heterogeneity accompanied by proven concordance between tissue and CTC genomic profiles [15]. Single-cell RNA sequencing (scRNAseq) recently demonstrated deep diversity by several different CTC subpopulations, including epithelial phenotype, epithelial-mesenchymal phenotype, mesenchymal phenotype, and stem phenotype [16] . Overall, this new perspective may identify subpopulations that lead to metastatic spread and treatment resistance, and thus cancer progression and recurrence [16, 17]. The challenge is to further investigate the phenotype of these tumors, and what is more difficult is their reliable amplification. This enables the emergence of new personalized tumor models that represent the patient’s tumor heterogeneity.
Live CTCs are rarely present in the bloodstream. Enriching this small number of live CTCs so that all these analyzes can be performed remains a huge challenge. Several methods have been used, most of which target the physical size, density, or expression of the tumor.
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