Model For Drug Screening Research Articles

Functional Drug Screening in the Era of Precision Medicine.

The focus of precision medicine is providing the right treatment to each unique patient. This scientific movement has incited monumental advances in oncology including the approval of effective, targeted agnostic therapies. Yet, precision oncology has focused largely on genomics in the treatment decision making process, and several recent clinical trials demonstrate that genomics is not the only variable to be considered. Drug screening in three dimensional (3D) models, including patient derived organoids, organs on a chip, xenografts, and 3D-bioprinted models provide a functional medicine perspective and necessary complement to genomic testing. In this review, we discuss the practicality of various 3D drug screening models and each model’s ability to capture the patient’s tumor microenvironment. We highlight the potential for enhancing precision medicine that personalized functional drug testing holds in combination with genomic testing and emerging mathematical models.

Cardiac ultrastructure inspired matrix induces advanced metabolic and functional maturation of differentiated human cardiomyocytes.

The vast potential of human induced pluripotent stem-cell-derived cardiomyocytes (hiPSC-CMs) in preclinical models of cardiac pathologies, precision medicine, and drug screening remains to be fully realized because hiPSC-CMs are immature without adult-like characteristics. Here, we present a method to accelerate hiPSC-CM maturation on a substrate, cardiac mimetic matrix (CMM), mimicking adult human heart matrix ligand chemistry, rigidity, and submicron ultrastructure, which synergistically mature hiPSC-CMs rapidly within 30days. hiPSC-CMs matured on CMM exhibit systemic transcriptomic maturation toward an adult heart state, are aligned with high strain energy, metabolically rely on oxidative phosphorylation and fatty acid oxidation, and display enhanced redox handling capability, efficient calcium handling, and electrophysiological features of ventricular myocytes. Endothelin-1-induced pathological hypertrophy is mitigated on CMM, highlighting the role of a native cardiac microenvironment in withstanding hypertrophy progression. CMM is a convenient model for accelerated development of ventricular myocytes manifesting highly specialized cardiac-specific functions.

Light‐induced hydrogels derived from poly(ethylene glycol) and acrylated methyl ricinoleate as biomaterials

AbstractHydrogels are hydrophilic crosslinked polymer networks that can absorb large amounts of water. They are used as biomaterials in numerous tissue engineering applications. Considering environmental awareness, the synthesis of biomaterials from renewable resources through green fabrication methods is essential. This study produces thermoresponsive hydrogels from a castor oil‐based monomer, acrylated methyl ricinoleate, and poly(ethylene glycol) via an environmentally friendly synthesis method. A photopolymerization technique is used with a very short reaction time. Characterization of the hydrogels is performed using thermogravimetric analysis, scanning electron microscopy, and Fourier‐transform infrared spectroscopy. Swelling and deswelling profiles are subsequently analyzed. A maximum equilibrium swelling degree of 271% is reached within 30 min. In vitro cytotoxicity assays of the hydrogels and the degradation products are performed to evaluate the biocompatibility. The hydrogels are biocompatible because the cell survival of all hydrogel samples and degradation products is greater than 100% and 85%, respectively. Consequently, the thermoresponsive hydrogels made from renewable raw materials in a green process offer interesting platforms for building biomaterials such as actuators for lab‐on‐a‐chip devices, microfluidics, drug delivery systems, preclinical drug screening models, and regenerative medicine.

Abstract 191: Establishment and characterization of patient-derived cancer cell models for pharmaceutical drug screening

Abstract Patient-derived cell (PDC) models are a tool that bridge the gap between conventional cancer lines that do not faithfully replicate clinical responses and costly and time-consuming patient derived xenograft (PDX) models. Imagen Therapeutics is establishing the world’s largest biobank of PDC models for both patient diagnostic (PredictRx) and biopharma drug development (PredictTx) studies that show a high degree of correlation between PDC and patient drug response. Through our research, we have established unique culture conditions that specifically select for cancer stem cells, and these have been characterized using both 2-D and 3-D short-term high content image (HCI)-based analysis. To better understand the genomic and phenotypic profiles of these models we analyzed Next Generation Sequencing data and phenotypic cell killing using a panel of FDA-approved agents. Our results showed that mRNA expression of several known cancer stem cell markers were upregulated in our PDCs in addition to multidrug resistance (MDR) genes. Drug resistance in the phenotypic assay was then correlated with stemness and MDR expression profiles. Finally, sequential PDCs (models developed from same patient at different stages in disease progression), in addition to PDCs developed from the same patient at multiple lesion sites, were compared in terms of genomic sequencing and drug responses. Together our data supports the premise that PDCs recapitulate the cancer stem cell population and are a useful platform with which to interrogate drug responses in a variety of different cancer types. Citation Format: Gareth J. Griffiths, Abbie Dodd, Olivia Nee, Tilly Thuringer, Subir Singh, Rachel Howard-Jones, Yuen Ngan Fan, Albert Bezman, Rachel Bell, Ido Sloma, Chris Noakes, Dominic I. James. Establishment and characterization of patient-derived cancer cell models for pharmaceutical drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 191.

Abstract 1890: Development of the drug efficacy testing in ex vivo 3D cultures (DETECT) platform and its application to functional precision medicine in ovarian cancer

Abstract Many ovarian cancer (OC) patients receiving chemotherapy relapse with acquired resistance, hence novel treatment allocation strategies are urgently needed. 3D cell culture models and organoid technologies have emerged as exciting tools to study cancer. Despite the advances, there is a need for protocols that can quickly generate patient-relevant 3D models for high-throughput drug screening and for diagnostic drug sensitivity testing. Ideally, such methods should be cost-effective, provide multi-parametric data to reveal distinct phenotypes, have single cell resolution to reveal heterogeneity and cell interactions, meet diagnostic timeframes and be miniaturized to allow the analysis of many drugs and combinations with small numbers of primary tumor cells. Here, we aimed to develop a robust technology for high-throughput ex vivo drug sensitivity testing for functional precision medicine using OC tissue and ascites samples (mostly high-grade serous carcinoma) from patients. We present a scalable drug screening platform: DETECT (Drug Efficacy Testing in Ex-vivo 3D Cultures), where tumor-derived patient cells can be rapidly screened within one week from sampling. Using this information patient-specific drug combinations are subsequently designed and tested with these data available within 9 days. This is much faster than with most screening protocols for organoids and PDC models where many weeks or months are required for establishment of the models and expansion of the required numbers of cells. We have currently tested the efficacy of 58 selected chemo- and targeted drugs in 5 doses using high content imaging and then a complete dose-matrix of 3 combinations at step 2. We report robust drug screening data in 15 out of 18 OC patient samples, which has resulted in a functional taxonomy of patient samples and a taxonomy of drugs based on their efficacy across patient samples. Some of the most common responses were seen for the BCL-XL inhibitor A-1331852 (9/15 patients), Topotecan (7), Dactinomycin (7), Omipalisib (6) and Omacetaxine (6). All but 2 drugs showed efficacy in at least one OC sample, suggesting heterogeneity and opportunities to make use of unique drug response patterns. Combination screening revealed that Carboplatin and A-1331852, a BCL-XL inhibitor, showed increased efficacy in 3 of the 5 tested patient samples. In conclusion, our 3D HT-drug testing assay DETECT with a combination screening capability could in the future be useful for guiding individualized treatment in a clinical setting as well as for identifying existing and emerging drugs and drug combinations for repurposing in OC. Citation Format: Emma Åkerlund, Greta Gudoityte, Elisabeth Moussaud Lamodiere, Joseph Carlson, Emelie Wallin, Josefin Fernebro, Olli Kallioniemi, Päivi Östling, Ulrika Joneborg, Brinton Seashore-Ludlow. Development of the drug efficacy testing in ex vivo 3D cultures (DETECT) platform and its application to functional precision medicine in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1890.

Pivotal role of micro-CT technology in setting up an optimized lung fibrosis mouse model for drug screening

Idiopathic pulmonary fibrosis (IPF) is a progressive disease with no curative pharmacological treatment. The most used animal model of IPF for anti-fibrotic drug screening is bleomycin (BLM)-induced lung fibrosis. However, several issues have been reported: the balance among disease resolution, an appropriate time window for therapeutic intervention and animal welfare remain critical aspects yet to be fully elucidated. In this study, C57Bl/6 male mice were treated with BLM via oropharyngeal aspiration (OA) following either double or triple administration. The fibrosis progression was longitudinally assessed by micro-CT every 7 days for 4 weeks after BLM administration. Quantitative micro-CT measurements highlighted that triple BLM administration was the ideal dose regimen to provoke sustained lung fibrosis up to 28 days. These results were corroborated with lung histology and Bronchoalveolar Lavage Fluid cells. We have developed a mouse model with prolonged lung fibrosis enabling three weeks of a curative therapeutic window for the screening of putative anti-fibrotic drugs. Moreover, we have demonstrated the pivotal role of longitudinal micro-CT imaging in reducing the number of animals required per experiment in which each animal can be its own control. This approach permits a valuable decrease in costs and time to develop disease animal models.

Abstract 1095: Establishment and characterization of a topotecan resistant lung cancer NCI H460/TPT10 cell line and a tumor xenograft model

Abstract While topotecan (TPT) is a chemotherapeutic drug in treating lung cancer, the development of TPT resistance in tumors reserves as a major obstacle to chemotherapeutic success. Therefore, a better understanding of the mechanisms of TPT resistance is critical. In this study, the first topotecan-resistant human non-small cell lung cancer (NSCLC) cell line, termed NCI-H460/TPT10, was established from the parental NCI-H460 cell line. NCI-H460/TPT10 cells exhibited a 394.7-fold resistance to TPT, and cross-resistance to SN-38, mitoxantrone, and doxorubicin, compared to parental NCI-H460 cells. Overexpression of ABCG2 localized on the cell membrane, but not ABCB1 or ABCC1, was found in NCI-H460/TPT10 cells, indicating that ABCG2 was likely to be involved in TPT-resistance. This was confirmed by the abolishment of drug resistance in NCI-H460/TPT10 cells after ABCG2 knockout. Moreover, the involvement of functional ABCG2 as a drug efflux pump conferring multidrug resistance (MDR) was indicated by low intracellular accumulation of TPT in NCI-H460/TPT10 cells, and the reversal effects by ABCG2 inhibitor Ko143 and cabozantinib. The NCI-H460/TPT10 and its parental cell line were further used to establish in vivo tumor xenograft mouse models, which verified their capability to serve as clinically relevant models for drug screening and the development of targeted strategies to overcome ABCG2-mediated MDR in NSCLC. Citation Format: Qiu-Xu Teng, Zi-Ning Lei, Wei Zhang, Ying-Fang Fan, Jing-Quan Wang, Chao-Yun Cai, Dong-Hua Yang, John Wurpel, Zhe-Sheng Chen. Establishment and characterization of a topotecan resistant lung cancer NCI H460/TPT10 cell line and a tumor xenograft model [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 1095.

Abstract 6039: Application of triple negative breast cancer patient-derived organoid and xenograft model for drug screening

Abstract Background: Breast cancer Patient Derived Organoids (PDOs) and patient-derived xenografts (PDXs) which recapitulate the tumor heterogeneity and molecular signatures of original tumor tissue, are useful tools to enable greater precision of both approved and experimental medicines through personalized therapeutic approaches. We focus on use of ex vivo PDOs, PDX-derived organoids (PDXOs), and vivo PDXs models for drug testing and personalized therapy. In this study, we investigated the oncogenic role and therapeutic potency of targeting WEE1 in breast cancer. Methods: Fresh surgical specimens of primary TNBC breast tumor (N=3) were used for PDOs. PDOs and PDX-derived organoids (PDXOs) were cultured in previously reported media compositions based on ECM hydrogel. PODs and PDXOs were subjected to drug screening for 22 anticancer drugs including chemoreagents and targeted drugs in the 3D HTS system. Drug sensitivity was tested in triplicate in different concentration ranges for 7 days. The IC50 for each drug was calculated by a sigmoidal dose-response curve, using the GraphPad Prism 9 program. For drug screening in PDXs models, PDOs were injected into the fourth mammary fat pads of NOD.Cg-Prkdcscid Il2rgtm1wjl/SzJ mice. WEE1 inhibitor (AZD1775) was administered by oral gavage (AZD1775, 30 mg/kg) for 3 weeks. Immunostaining of ER, PR, Her2, CK5 and Ki67 was performed in PODs, PODXs and PDX tumor tissues. Results: All PDOs, PDXs, and PDXOs were ER-, PR- and HER2- negative. PDOs and PDXOs (172T, 185T, 207T) showed differential response to 22 anticancer drugs. In PDOs and PDXOs models, 172T and 207T PDOs and PDXOs are highly sensitive to AZD1775, but 185T PDOs and PDXOs less sensitive or never respond to treatment with AZD1775 (10 µM). In PDX models, treatment with AZ1775 decreased tumor growth (P<0.0079) as well as distance metastasis in 207T-PDX mice, whereas in 172T- and 185T-PDX mice, the tumor growth and metastasis were not reduced compared with the vehicle group. Conclusion: Consequently, PDOs, PDXOs and PDXs recapitulated immunohistological signatures of original tumor tissues. The results of WEE1 inhibitor response between PDOs, PDXOs and PDXs were consistent. This study may provide a preclinical tool to screen drug responses to standard-of-care and newly identified drugs. Citation Format: Seungyeon Ryu, Hoe Suk Kim, Jung Eun KIM, So-Hyun Yoon, Sangeun Lee, Moonjou Baek, Han-Byoel Lee, Dong Woo Lee, Bosung Ku, Wonshik Han. Application of triple negative breast cancer patient-derived organoid and xenograft model for drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6039.

Dnah9 mutant mice and organoid models recapitulate the clinical features of patients with PCD and provide an excellent platform for drug screening

Primary cilia dyskinesia (PCD) is a rare genetic disease caused by ciliary structural or functional defects. It causes severe outcomes in patients, including recurrent upper and lower airway infections, progressive lung failure, and randomization of heterotaxy. To date, although 50 genes have been shown to be responsible for PCD, the etiology remains elusive. Meanwhile, owing to the lack of a model mimicking the pathogenesis that can be used as a drug screening platform, thereby slowing the development of related therapies. In the current study, we identified compound mutation of DNAH9 in a patient with PCD with the following clinical features: recurrent respiratory tract infections, low lung function, and ultrastructural defects of the outer dynein arms (ODAs). Bioinformatic analysis, structure simulation assay, and western blot analysis showed that the mutations affected the structure and expression of DNAH9 protein. Dnah9 knock-down (KD) mice recapitulated the patient phenotypes, including low lung function, mucin accumulation, and increased immune cell infiltration. Immunostaining, western blot, and co-immunoprecipitation analyses were performed to clarify that DNAH9 interacted with CCDC114/GAS8 and diminished their protein levels. Furthermore, we constructed an airway organoid of Dnah9 KD mice and discovered that it could mimic the key features of the PCD phenotypes. We then used organoid as a drug screening model to identify mitochondrial-targeting drugs that can partially elevate cilia beating in Dnah9 KD organoid. Collectively, our results demonstrated that Dnah9 KD mice and an organoid model can recapture the clinical features of patients with PCD and provide an excellent drug screening platform for human ciliopathies.

Generation of an erythroid progenitor-derived iPSC line, VRISGi002-A, from a healthy 27-year-old Vietnamese donor under a feeder-free system

We have established the footprint-free Vietnamese human induced pluripotent stem cell (hiPSC) line, VRISGi002-A, from CD71 + CD235a + erythroid progenitor cells (EPCs) of a 27-year-old healthy donor. The EPCs were enriched from isolated peripheral blood and reprogrammed using Sendai viruses which carried the reprogramming factors c-MYC, SOX2, KLF4, and OCT4 under a feeder-free culture system. The established VRISGi002-A cell line expressed typical pluripotency markers, displayed a normal karyotype, and demonstrated the potential to differentiate into the three germ layers. This hiPSC line could serve as a Vietnamese healthy control model for physiological processes and drug screening.

Vascularization of Patient-Derived Tumoroid from Non-Small-Cell Lung Cancer and Its Microenvironment.

Patient-derived tumoroid (PDT) has been developed and used for anti-drug screening in the last decade. As compared to other existing drug screening models, a PDT-based in vitro 3D cell culture model could preserve the histological and mutational characteristics of their corresponding tumors and mimic the tumor microenvironment. However, few studies have been carried out to improve the microvascular network connecting the PDT and its surrounding microenvironment, knowing that poor tumor-selective drug transport and delivery is one of the major reasons for both the failure of anti-cancer drug screens and resistance in clinical treatment. In this study, we formed vascularized PDTs in six days using multiple cell types which maintain the histopathological features of the original cancer tissue. Furthermore, our results demonstrated a vascular network connecting PDT and its surrounding microenvironment. This fast and promising PDT model opens new perspectives for personalized medicine: this model could easily be used to test all therapeutic treatments and could be connected with a microfluidic device for more accurate drug screening.

PD43-08 DEVELOPMENT OF PATIENT-DERIVED KIDNEY CANCER ORGANOID; APPLICATION TO DRUG SCREENING MODELS AND ITS CHALLENGES

You have accessJournal of UrologyCME1 May 2022PD43-08 DEVELOPMENT OF PATIENT-DERIVED KIDNEY CANCER ORGANOID; APPLICATION TO DRUG SCREENING MODELS AND ITS CHALLENGES Akira Kazama, Vladimir Bilim, and Yoshihiko Tomita Akira KazamaAkira Kazama More articles by this author , Vladimir BilimVladimir Bilim More articles by this author , and Yoshihiko TomitaYoshihiko Tomita More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002604.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The selection of effective therapeutic agents is critical to improve the life prognosis of patients with renal cell carcinoma (RCC). In this study, we established 3-dimensional (3D) tumor organoids from freshly resected RCC tumors and tested tyrosine kinase inhibitors (TKIs), targeted therapeutics for metastatic RCC, as a drug testing model for development of potential personalized therapy for RCC patients. METHODS: Surgical tumor specimens were obtained from 20 patients with RCC. 3D tumor organoids were developed ex vivo from freshly resected RCC tumors and their histopathological and molecular characteristics were evaluated using histological staining and whole exome sequencing (WES). Using cell viability assay, we determined the therapeutic efficacy of TKIs in RCC tumor organoids. RESULTS: We found that RCC tumor organoids recapitulate the histological architecture of the primary tumor. Using WES, we identified strong concordance of primary tumors and corresponding tumor organoids at genetic level. We demonstrated that RCC tumor organoids respond differently to targeted therapeutics showing patient-specific sensitivity to the treatment. CONCLUSIONS: Our results identify concordance of genetic aberrations in primary RCC and corresponding tumor organoids, and demonstrate that tumor organoid models allow ex vivo drug testing of tumor samples obtained from RCC patients to predict a potential clinical response to targeted therapeutics. Source of Funding: The present study was supported by a research grant from the Department of Urology, Division of Molecular Oncology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan (grant no. CH29017; August, 2018) © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e704 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Akira Kazama More articles by this author Vladimir Bilim More articles by this author Yoshihiko Tomita More articles by this author Expand All Advertisement PDF DownloadLoading ...

Contractility analysis of human engineered 3D heart tissues by an automatic tracking technique using a standalone application.

The use of Engineered Heart Tissues (EHT) as in vitro model for disease modeling and drug screening has increased, as they provide important insight into the genetic mechanisms, cardiac toxicity or drug responses. Consequently, this has highlighted the need for a standardized, unbiased, robust and automatic way to analyze hallmark physiological features of EHTs. In this study we described and validated a standalone application to analyze physiological features of EHTs in an automatic, robust, and unbiased way, using low computational time. The standalone application “EHT Analysis” contains two analysis modes (automatic and manual) to analyzes the contractile properties and the contraction kinetics of EHTs from high speed bright field videos. As output data, the graphs of displacement, contraction force and contraction kinetics per file will be generated together with the raw data. Additionally, it also generates a summary file containing all the data from the analyzed files, which facilitates and speeds up the post analysis. From our study we highlight the importance of analyzing the axial stress which is the force per surface area (μN/mm2). This allows to have a readout overtime of tissue compaction, axial stress and leave the option to calculate at the end point of an experiment the physiological cross-section area (PSCA). We demonstrated the utility of this tool by analyzing contractile properties and compaction over time of EHTs made out of a double reporter human pluripotent stem cell (hPSC) line (NKX2.5EGFP/+-COUP-TFIImCherry/+) and different ratios of human adult cardiac fibroblasts (HCF). Our standalone application “EHT Analysis” can be applied for different studies where the physiological features of EHTs needs to be analyzed under the effect of a drug compound or in a disease model.

A high-throughput biomimetic bone-on-a-chip platform with artificial intelligence-assisted image analysis for osteoporosis drug testing.

Although numerous organ-on-a-chips have been developed, bone-on-a-chip platforms have rarely been reported because of the high complexity of the bone microenvironment. With an increase in the elderly population, a high-risk group for bone-related diseases such as osteoporosis, it is essential to develop a precise bone-mimicking model for efficient drug screening and accurate evaluation in preclinical studies. Here, we developed a high-throughput biomimetic bone-on-a-chip platform combined with an artificial intelligence (AI)-based image analysis system. To recapitulate the key aspects of natural bone microenvironment, mouse osteocytes (IDG-SW3) and osteoblasts (MC3T3-E1) were cocultured within the osteoblast-derived decellularized extracellular matrix (OB-dECM) built in a well plate-based three-dimensional gel unit. This platform spatiotemporally and configurationally mimics the characteristics of the structural bone unit, known as the osteon. Combinations of native and bioactive ingredients obtained from the OB-dECM and coculture of two types of bone cells synergistically enhanced osteogenic functions such as osteocyte differentiation and osteoblast maturation. This platform provides a uniform and transparent imaging window that facilitates the observation of cell-cell interactions and features high-throughput bone units in a well plate that is compatible with a high-content screening system, enabling fast and easy drug tests. The drug efficacy of anti-SOST antibody, which is a newly developed osteoporosis drug for bone formation, was tested via β-catenin translocation analysis, and the performance of the platform was evaluated using AI-based deep learning analysis. This platform could be a cutting-edge translational tool for bone-related diseases and an efficient alternative to bone models for the development of promising drugs.

Multi-omics analysis based on 3D-bioprinted models innovates therapeutic target discovery of osteosarcoma.

Current in vitro models for osteosarcoma investigation and drug screening, including two-dimensional (2D) cell culture and tumour spheroids (i.e. cancer stem-like cells), lack extracellular matrix (ECM). Therefore, results from traditional models may not reflect real pathological processes in genuine osteosarcoma histological structures. Here, we report a three-dimensional (3D) bioprinted osteosarcoma model (3DBPO) that contains osteosarcoma cells and shrouding ECM analogue in a 3D frame. Photo-crosslinkable bioinks composed of gelatine methacrylamide and hyaluronic acid methacrylate mimicked tumour ECM. We performed multi-omics analysis, including transcriptomics and DNA methylomics, to determine differences between the 3DBPO model and traditional models. Compared with 2D models and tumour spheroids, our 3DBPO model showed significant changes in cell cycle, metabolism, adherens junctions, and other pathways associated with epigenetic regulation. The 3DBPO model was more sensitive to therapies targeted to the autophagy pathway. We showed that simulating ECM yielded different osteosarcoma cell metabolic characteristics and drug sensitivity in the 3DBPO model compared with classical models. We suggest 3D printed osteosarcoma models can be used in osteosarcoma fundamental and translational research, which may contribute to novel therapeutic strategy discovery.

Fundamental signaling pathways for glioblastoma drug resistance and developing robust organoid models for drug discovery

This article presents a review of the current literature on the molecular mechanisms of treatment resistance in glioblastoma. As mounting research continues to explore novel methods of treating glioblastoma, from using organoid models for drug screening to developing novel cellular therapies, it is critical to understand the fundamental molecular landscape that makes glioblastoma difficult to treat. This review explores the means of chemoresistance to the conventional chemotherapy temozolomide. Consideration of DNA repair pathways, p53-mediated apoptosis and autophagy, convergent proliferation pathways, and epigenetic mechanisms demonstrate avenues for the development of sophisticated drug targets and combination treatments. Ultimately, this article highlights each of these mechanisms and presents referential material for future endeavors in organoid-based drug screening.

Use of conditional reprogramming cell, patient derived xenograft and organoid for drug screening for individualized prostate cancer therapy: Current and future perspectives (Review).

Prostate cancer mortality is ranked second among all cancer mortalities in men worldwide. There is a great need for a method of efficient drug screening for precision therapy, especially for patients with existing drug‑resistant prostate cancer. Based on the concept of bacterial cell culture and drug sensitivity testing, the traditional approach of cancer drug screening is inadequate. The current and more innovative use of cancer cell culture and invivo tumor models in drug screening for potential individualization of anti‑cancer therapy is reviewed and discussed in the present review. An ideal screening model would have the ability to identify drug activity for the targeted cells resembling what would have occurred in the invivo environment. Based on this principle, three available cell culture/tumor screening models for prostate cancer are reviewed and considered. The culture conditions, advantages and disadvantages for each model together with ideas to best utilize these models are discussed. The first screening model uses conditional reprogramed cells derived from patient cancer cells. Although these cells are convenient to grow and use, they are likely to have different markers and characteristics from original tumor cells and thus not likely to be informative. The second model employs patient derived xenograft (PDX) which resembles an invivo approach, but its main disadvantages are that it cannot be easily genetically modified and it is not suitable for high‑throughput drug screening. Finally, high‑throughput screening is more feasible with tumor organoids grown from patient cancer cells. The last system still needs a large number of tumor cells. It lacks insitu blood vessels, immune cells and the extracellular matrix. Based on these current models, future establishment of an organoid data bank would allow the selection of a specific organoid resembling that of an individual's prostate cancer and used for screening of suitable anticancer drugs. This can be further confirmed using the PDX model. Thus, this combined organoid‑PDX approach is expected to be able to provide the drug sensitivity testing approach for individualization of prostate cancer therapy in the near future.

3D Printing in Solid Dosage Forms and Organ-on-Chip Applications.

3D printing (3DP) can serve not only as an excellent platform for producing solid dosage forms tailored to individualized dosing regimens but can also be used as a tool for creating a suitable 3D model for drug screening, sensing, testing and organ-on-chip applications. Several new technologies have been developed to convert the conventional dosing regimen into personalized medicine for the past decade. With the approval of Spritam, the first pharmaceutical formulation produced by 3DP technology, this technology has caught the attention of pharmaceutical researchers worldwide. Consistent efforts are being made to improvise the process and mitigate other shortcomings such as restricted excipient choice, time constraints, industrial production constraints, and overall cost. The objective of this review is to provide an overview of the 3DP process, its types, types of material used, and the pros and cons of each technique in the application of not only creating solid dosage forms but also producing a 3D model for sensing, testing, and screening of the substances. The application of producing a model for the biosensing and screening of drugs besides the creation of the drug itself, offers a complete loop of application for 3DP in pharmaceutics.

Culture of patient-derived multicellular clusters in suspended hydrogel capsules for pre-clinical personalized drug screening.

A personalized medication regimen provides precise treatment for an individual and can be guided by pre-clinical drug screening. The economical and high-efficiency simulation of the liver tumor microenvironment (TME) in a drug-screening model has high value yet challenging to accomplish. Herein, we propose a simulation of the liver TME with suspended alginate-gelatin hydrogel capsules encapsulating patient-derived liver tumor multicellular clusters, and the culture of patient-derived tumor organoids(PDTOs) for personalized pre-clinical drug screening. The hydrogel capsule offers a 3D matrix environment with mechanical and biological properties similar to those of the liver in vivo. As a result, 18 of the 28 patient-derived multicellular clusters were successfully cultured as PDTOs. These PDTOs, along with hepatocyte growth factor (HGF) of non-cellular components, preserve stromal cells, including cancer-associated fibroblasts (CAFs) and vascular endothelial cells (VECs). They also maintain stable expression of molecular markers and tumor heterogeneity similar to those of the original liver tumors. Drugs, including cabazitaxel, oxaliplatin, and sorafenib, were tested in PDTOs. The sensitivity of PDTOs to these drugs differs between individuals. The sensitivity of one PDTO to oxaliplatin was validated using magnetic resonance imaging (MRI) and biochemical tests after oxaliplatin clinical treatment of the corresponding patient. Therefore, this approach is promising for economical, accurate, and high-throughput drug screening for personalized treatment.

Advanced In Vitro Lung Models for Drug and Toxicity Screening: The Promising Role of Induced Pluripotent Stem Cells (Adv. Biology 2/2022)

Artificial Lung Models In article 2101139, Costa and co-workers use of biomanufacturing (electrospinning/3D-printing) and lung-on-chip technology for the development of artificial lung models. As such, microfluidic channels and 3D-printed fibers are arranged in order to represent the respiratory tree and pulmonary silhouette, simulating larger bronchi dividing into consecutively smaller branches and, finally, the alveoli.