Drug Testing System Research Articles

Evaluation of a Point of Care Urine Drug Test System for Application in Outpatient Pain Management Clinics

Abstract Introduction/Objective Most states require testing for illicit and controlled substances in ambulatory, non-terminal pain management populations to ensure prescription compliance. Point of care (POC) urine drug testing (UDT) is a widely adopted testing method that assists with clinical decision making within minutes. However, these immunoassays may lack the sensitivity and specificity needed for potent medications such as fentanyl. This study compares the performance characteristics of a POC UDT to an in-house, high throughput, automated chemistry analyzer and the reference method, gas chromatography-mass spectrometry (GC-MS). Providers should be aware of the limitations of each methodology given the rise of synthetic opioid misuse, and the need to quantify multiple substances and metabolites within the same drug class. Methods/Case Report 100 residual deidentified patient urine samples from an ambulatory pain management population were analyzed using GC-MS, Profile®-V MEDTOX Scan® POCT device (Burlington, NC), and a Beckman Coulter AU480® Chemistry Analyzer using EMIT® II Plus reagents (Beckman Coulter; Brea, CA). Drug classes on the Profile®-V included: amphetamines, barbiturates, benzodiazepines, buprenorphine, cocaine metabolite, methamphetamine, methadone, opiates, oxycodone, cannabinoids and phencyclidine. Drug classes on the AU480® included the same except without a methamphetamine-specific immunoassay. Results (if a Case Study enter NA) The results from GC-MS showed that 99% (99/100) of the samples were positive for one or more of the drug classes. Overall, the AU480® was 90% sensitive and 98% specific while the MEDTOX Scan® was 87% sensitive and 98% specific. Both devices exhibited variable performance across drug classes. Neither the AU480® nor the MEDTOX Scan® detected fentanyl in five specimens. Conclusion POCT-UDS systems like the Profile®-V MEDTOX Scan® can provide good accuracy and may offer a time and cost benefit when compared to higher throughput analyzers like the AU-480®. Due to the low reference values of immunoassays, definitive testing such as GC-MS is recommended for use in the pain management population.

의료용 마약류 오남용에 대한 대응 방안

The social problems caused by the misuse of medical drugs are increasing. Therefore, countermeasures to reduce the misuse of medical drugs are needed. First, as a way to control the supply of medical drugs, it is necessary to increase the number of medical drug monitoring personnel, link the NIMS with the drug safety service system, revise the Act so that it can be reported immediately after the prescription of the drug for medical use, establish a legal basis for reporting the plan to dispose of medical drugs to the authorities when medical institutions and pharmacies are closed, establish a legal basis for a total ban on business if the reporting of drug handling details is violated, improve medical drug return regulations under the Narcotics Control Act, establish objective standards for re-issuance of medical licenses, strengthen medical drug education for doctors and pharmacists, restrict medical drug prescription qualifications, improve the system to handle medical drugs only at high-level hospitals, and introduce a system to require identification through the introduction of a fingerprint search system when prescribing medical drugs. Second, as a way to curb the demand for misuse of medical drugs, the following are required: The establishment of an infrastructure for collecting and disposing of medical drugs at home, establishing an investigative organization specializing in medical drugs, strengthening the iInternet monitoring system, education and promotion of the harmfulness of medical drugs, introducing compulsory treatment and education order systems, introducing a drug treatment and testing system, and integrating medical drug offenders for treatment purposes.

Inflammation-related molecular signatures involved in the anticancer activities of brigatinib as well as the prognosis of EML4-ALK lung adenocarcinoma patient.

Although ALK tyrosine kinase inhibitors (ALK-TKIs) have shown remarkable benefits in EML4-ALK positive NSCLC patients compared to conventional chemotherapy, the optimal sequence of ALK-TKIs treatment remains unclear due to the emergence of primary and acquired resistance and the lack of potential prognostic biomarkers. In this study, we systematically explored the validity of sequential ALK inhibitors (alectinib, lorlatinib, crizotinib, ceritinib and brigatinib) for a heavy-treated patient with EML4-ALK fusion via developing an in vitro and in vivo drug testing system based on patient-derived models. Based on the patient-derived models and clinical responses of the patient, we found that crizotinib might inhibit proliferation of EML4-ALK positive tumors resistant to alectinib and lorlatinib. In addition, NSCLC patients harboring the G1269A mutation, which was identified in alectinib, lorlatinib and crizotinib-resistant NSCLC, showed responsiveness to brigatinib and ceritinib. Transcriptomic analysis revealed that brigatinib suppressed the activation of multiple inflammatory signaling pathways, potentially contributing to its anti-tumor activity. Moreover, we constructed a prognostic model based on the expression of IL6, CXCL1, and CXCL5, providing novel perspectives for predicting prognosis in EML4-ALK positive NSCLC patients. In summary, our results delineate clinical responses of sequential ALK-TKIs treatments and provide insights into the mechanisms underlying the superior effects of brigatinib in patients harboring ALKG1269A mutation and resistant towards alectinib, lorlatinib and crizotinib. The molecular signatures model based on the combination of IL6, CXCL1 and CXCL5 has the potential to predict prognosis of EML4-ALK positive NSCLC patients.

Bioreactor Culture to Create Adipose Tissue from Human Mesenchymal Stromal Cells.

Adipose tissue is a complex and multifaceted endocrine organ located throughout the body. The dysfunction of adipose tissue is known to induce a wide variety of comorbidities that can negatively impact one's health and quality of life. In addition to behavioral changes, drugs that target dysfunctional adipose tissue to treat associated diseases are clinically needed. Regarding drug-testing platforms, animal models are the most popular models, limited by known differences from humans in genetics and physiology. Two-dimensional and static three-dimensional (3D) cell cultures are also used. Still, these in vitro models with static culture fail to recapitulate the phenotype and function of adipocytes seen in vivo. To combat this, our lab has developed an adipose tissue microphysiological system. A perfusion bioreactor with dual-flow chambers is 3D printed, which enables individualized top and bottom medium flows after adipose tissues are inserted as a barrier. Human progenitor cells, such as human mesenchymal stem cells, are embedded within a gelatin scaffold and in situ adipogenic differentiation within the bioreactor. Medium flow is established via a syringe pump system, allowing in vivo-like conditions to be maintained. The novel bioreactor-cultured adipose tissues represent a versatile disease modeling and drug-testing system. Here, we present the step-by-step methods to generate the bioreactors and adipose tissues. We also show the process of collecting and analyzing samples. In addition, we highlight the critical steps that require particular attention in notes.

Scaffold-based spheroid models of glioblastoma multiforme and its use in drug screening.

Among several types of brain cancers, glioblastoma multiforme (GBM) is a terminal and aggressive disease with a median survival of 15 months despite the most intensive surgery and chemotherapy. Preclinical models that accurately reproduce the tumor microenvironment are vital for developing new therapeutic alternatives. Understanding the complicated interactions between cells and their surroundings is essential to comprehend the tumor's microenvironment, however the monolayer cell culture approach falls short. Numerous approaches are used to develop GBM cells into tumor spheroids, while scaffold-based spheroids provides the opportunity to investigate the synergies between cells as well as cells and the matrix. This review summarizes the development of various scaffold-based GBM spheroid models and the prospective for their use as drug testing systems.

Effect of PD-1 inhibition on macrophage population in human tissue cultures.

463 Background: Moderate response rates of PD-1 inhibition in gastric and esophagogastric junction cancers (GC/EGJC) urge for reliable biomarkers. To further improve treatment outcomes, patients need targeted therapy based on clinically relevant biomarkers and predictive test systems for assessing individual drug susceptibility. We have previously established patient derived tissue cultures (PDTC) as a comprehensive ex-vivo drug testing system for GC/EGJC patients. Methods: The standardized PDTC) model was developed using tissue from surgical resection specimens to investigate response to the PD-1 inhibitor Nivolumab and the macrophage depriving bisphosphonates clodronate and zoledronic acid. Resident macrophage population (CD68, CD163), tumor proliferation (Ki67) and apoptosis (cPARP), as well as bulk gene expression data (nanoString technology) were analysed after 72 hours of PD-1 inhibition. Additionally, supernatants were collected, and cytokine concentrations are measured based on Luminex technology. Results: Macrophage depletion could lead to enhanced anti-tumoral effects in PDTCs after PD-1 inhibition. Gene signatures for Interferon γ-mediated inflammation and myeloid cell activity were identified and found to be relevant indicators for tissue response upon immune checkpoint inhibition. We observed alterations in cytokine and soluble surface marker concentrations, such as IL-8, IL-17A and IL-23 or PD-L2, LAG3 and CTLA4. Conclusions: Yielding an organotypic cell composition, we were able to demonstrate individual response patterns and immunological characteristics to identify responsiveness to PD-1 inhibition in GC and EGJC tissue. PDTCs are potent to define biological marker sets ex vivo and to differentiate and predict clinical patient response as well as their inherent immune cell capacity and plasticity.

Microfluidically-generated Encapsulated Spheroids (μ-GELS): An All-Aqueous Droplet Microfluidics Platform for Multicellular Spheroids Generation.

Spheroids are three-dimensional clusters of cells that serve as in vitro tumor models to recapitulate in vivo morphology. A limitation of many existing on-chip platforms for spheroid formation is the use of cytotoxic organic solvents as the continuous phase in droplet generation processes. All-aqueous methods do not contain cytotoxic organic solvents but have so far been unable to achieve complete hydrogel gelation on chip. Here, we describe an enhanced droplet microfluidic platform that achieves on-chip gelation of all-aqueous hydrogel multicellular spheroids (MCSs). Specifically, we generate dextran-alginate droplets containing MCF-7 breast cancer cells, surrounded by polyethylene glycol, at a flow-focusing junction. Droplets then travel to a second flow-focusing junction where they interact with calcium chloride and gel on chip to form hydrogel MCSs. On-chip gelation of the MCSs is possible here because of an embedded capillary at the second junction that delays the droplet gelation, which prevents channel clogging problems that would otherwise exist. In drug-free experiments, we demonstrate that MCSs remain viable for 6 days. We also confirm the applicability of this system for cancer drug testing by observing that dose-dependent cell death is achievable using doxorubicin.

LUHMES Cells: Phenotype Refinement and Development of an MPP+-Based Test System for Screening Antiparkinsonian Drugs

The low effectiveness of symptomatic pharmacotherapy for Parkinson's disease (PD), which compensates for dopamine (DA) deficiency under degeneration of nigrostriatal dopaminergic (DAergic) neurons, could apparently be improved with neuroprotective therapy, which slows down neurodegeneration and PD progression. For this, it is necessary to have a DAergic cell line for the development of a PD model to screen neuroprotectors. We used immortalized human embryonic mesencephalon LUHMES cells (LCs) differentiated into DAergic neurons. The aim of this study was to characterize the phenotype of differentiated LCs and develop an 1-methyl-4-phenylpyridinium iodide (MPP+)-based test system for screening neuroprotectors. Using polymerase chain reaction (PCR) and immunocytochemistry, it has been shown that all differentiated LCs express genes and synthesize proteins characteristic of all neurons (microtubule-associated protein 2, bIII-tubulin, synaptotagmin 1) and specifically of DAergic neurons (tyrosine hydroxylase, aromatic L-amino acid decarboxylase, DA transporter, vesicular monoamine transporter 2). Furthermore, LCs are able to produce a small amount of DA, but under special conditions. To assess the mechanisms of neurodegeneration and neuroplasticity under the influence of toxins and antiparkinsonian drugs, including neuroprotectors, we have developed an LCs-based MPP+ PD model and proposed an original panel of markers for testing functional and structural cell disorders.

Analysis of polymeric nanoparticle properties for siRNA/DNA delivery in a tumor xenograft tissue slice air-liquid interface model.

Classical two-dimensional (2D) cell culture as a drug or nanoparticle test system only poorly recapitulates in vivo conditions. Animal studies are costly, ethically controversial, and preclude large-scale testing. We established a three-dimensional (3D) tissue slice air-liquid interface (ALI) culture model for nanoparticle testing. We developed an optimized procedure for the reproducible generation of large sets of tissue slices from tumor xenografts that retain their tissue architecture. When used for the analysis of nanoparticles based on chemically modified polyethylenimines (PEIs) to deliver siRNA or DNA, differences in transfection efficacy and cytotoxicity between nanoparticles were observed more clearly than in 2D cell culture. While nanoparticle efficacies between cell culture and the tissue slice model overall correlated, the tissue slice model also identified particularly suitable candidates whose efficacy was underestimated in 2D cell culture and had already been shown in previous in vivo studies. The ex vivo 3D tissue slice ALI culture model is a powerful system that allows the effective evaluation of biological nanoparticle efficacy and biocompatibility in an intact tissue environment. It is comparably inexpensive, time-saving, and follows the 3R principle, while allowing the identification of critical nanoparticle properties and optimal candidates for in vivo applications.

A Bone Marrow Mimetic 3D Co-Culture Model for Ex Vivo drug Testing of Down Syndrome-Myeloid Leukemia Cells

Children with Down syndrome are at elevated risk (10-20-fold) for the development of leukemia. The DS-myeloid leukemia (DS-ML) treatment relies largely on low-dose cytarabine (Ara-C). Despite the favorable outcome, there is 20% mortality rate associated with DS-ML. There is no targeted therapy so far for the patients who fail to respond to Ara-C treatment. Majority of clinical trials exploring investigational drugs for AML exclude DS patients because of differential drug sensitivity. Patient-derived xenograft models of DS-ML are limited, because of low sample availability and low success of engraftment. Therefore, an ex vivo drug testing model for DS-ML that can avoid the pitfalls of 2D culture system is a powerful tool to test novel therapies and understand the drug response mechanisms. To overcome the shortfalls of the previous in vitro drug testing systems for AML, we developed a 3D model that facilitates efficient interaction between leukemia cells with bone marrow mesenchymal stem cells (BM-MSCs) in a 3D microenvironment mimicking the extracellular matrix (ECM) components and modulus of BM. We generated pediatric BM-MSC lines from donor BM aspirates by their ability to adhere to tissue culture plastic. The immunophenotype analysis revealed that BM-MSC lack the expression of CD34 and CD45 but express CD90, CD44, CD105, CD73, CD166 and CD146. MSCs retained the potential of differentiation into the osteogenic, chondrogenic and adipogenic lineages. In addition to the cellular component, the BM microenvironment consists of various ECM proteins that contribute to the mechanics of the BM, and have profound effect on cellular phenotype and metabolism. PEG provides a blank slate for analyzing the peptide specific cellular responses, and can be further tuned to specify 3D architecture. The hydrogel-based 3D gel system that we developed for the generation of 3D model for transient abnormal myelopoiesis (Sidhu et al., Biomater Sci, 9:6266, 2021) was further tuned by the incorporation of BM mimetic peptides with composition and ratio determined by a proteomics study using human BM (Jansen et al., bioRxiv, 2018). To develop a BM model for drug testing, PEG gels containing BM-ECM mimicking peptides were used to encapsulate CMK cells alone and in co-culture with BM-MSCs. CMK cells treated with 0.5µM of Ara-C showed higher viability in co-culture (73% ± 12%) when compared to monoculture (52% ± 5%) (Figure 1A; bars 3 & 4; p*= 0.012). Similar to Ara-C, treatment with 0.75µM Azacitidine (Aza, a DNA hypomethylating agent) had a significantly higher viability in co-culture (90% ± 5%) compared to monoculture (74% ± 7%) (Figure 1A; bars 5 & 6; p*= 0.031). The chemoprotection of BM-MSCs against individual drug (Ara-C and Aza) was overridden by the sequential therapy with 0.75µM Aza for 24h, followed by 48h treatment with 0.5µM Ara-C under BM mimetic conditions. Compared to Ara-C alone treatment, cell viability was significantly lower in sequential therapy, with 39% (Figure 1A; bars 4 & 7; p#< 0.01) lower cell viability. Similarly, the sequential therapy reduced the cell viability by 56% when compared to Aza alone treatment (Figure 1C; Bars 6 and 7; p$< 0.01). This implies that Aza treatment can sensitize DS-ML cells to Ara-C treatment to disrupt the chemoprotective environment provided by BM-MSCs. In vitro megakaryocytic DS-ML cells harboring GATA1 and STAG2 mutations (T21-G1-S40) showing DS-ML characteristics were generated as described previously (Barwe et al., Cells, 11:628, 2022). The effect of Aza sensitization was tested in megakaryocytic lineage expanded T21-G1-S40 cells. Similar to CMK, the sequential treatment of T21-G1-S40 megakaryocytic cells show reduced cell viability (62% ± 6%; Figure 1B, bar 4) than the individual treatment with Ara-C (73% ± 7%; Figure 1B, bar2) and Aza (92% ± 6%; Figure 1B, bar 3). In summary, we established a synthetic 3D BM mimetic ex-vivo model consisting of the ECM components, mechanics and one of the key cellular components (MSCs) of BM microenvironment. The pediatric BM-MSCs induce chemoprotection to cytotoxic drugs in DS-ML cells. This chemoprotection can be overridden by the use of epigenetic modifier azacitidine. This study provides proof of principle for establishing a BM mimicking ex vivo model that can be utilized for high throughput drug screening. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

A Tumor‐Organoid‐Based Precision Medicine Platform for the Prediction of Drug Sensitivity of Colorectal Cancer

AbstractInvestigation of clinical actionable targets is crucial for the individual treatment of patients with advanced colorectal cancer (CRC). A complete platform integrating the molecular profiling and prediction of drug reactivity is currently scarce. An integrated precision medicine approach which combines high‐throughput sequencing platform with microfluidic organoid drug testing system is established. Capture sequencing identifies 6 drug targetable genes and 13 candidate drugs for a stage III colorectal cancer patient. The drug sensitivity is confirmed by using patient‐derived organoids (PDOs) model and microfluidic drug testing system. With this integrated precision medicine approach, the optimal drugs including Afatinib, Dacomitinib, and Osimertinib are successfully identified. The platform provides a practical method to guide the clinical therapies for colorectal cancer patients.

3D printed human organoids: High throughput system for drug screening and testing in current COVID-19 pandemic.

In the current pandemic, scenario the world is facing a huge shortage of effective drugs and other prophylactic medicine to treat patients which created havoc in several countries with poor resources. With limited demand and supply of effective drugs, researchers rushed to repurpose the existing approved drugs for the treatment of COVID-19. The process of drug screening and testing is very costly and requires several steps for validation and treatment efficacy evaluation ranging from in-vitro to in-vivo setups. After these steps, a clinical trial is mandatory for the evaluation of treatment efficacy and side effects in humans. These processes enhance the overall cost and sometimes the lead molecule show adverse effects in humans and the trial ends up in the final stages. Recently with the advent of three-dimensional (3D) organoid culture which mimics the human tissue exactly the process of drug screening and testing can be done in a faster and cost-effective manner. Further 3D organoids prepared from stems cells taken from individuals can be beneficial for personalized drug therapy which could save millions of lives. This review discussed approaches and techniques for the synthesis of 3D-printed human organoids for drug screening. The key findings of the usage of organoids for personalized medicine for the treatment of COVID-19 have been discussed. In the end, the key challenges for the wide applicability of human organoids for drug screening with prospects of future orientation have been included.

Accurate Evaluation of Hepatocyte Metabolisms on a Noble Oxygen-Permeable Material With Low Sorption Characteristics.

In the pharmaceutical industry, primary cultured hepatocytes is a standard tool used to assess hepatic metabolisms and toxicity in vitro. Drawbacks, however, include their functional deterioration upon isolation, mostly due to the lack of a physiological environment. Polydimethylsiloxane (PDMS) has been reported to improve the function of isolated hepatocytes by its high oxygen permeability when used as a material of microphysiological systems (MPS). However, its high chemical sorption property has impeded its practical use in drug development. In this study, we evaluated a new culture material, 4-polymethyl-1-pentene polymer (PMP), in comparison with PDMS and conventional tissue culture polystyrene (TCPS). First, we confirmed the high oxygen permeability and low sorption of PMP, and these properties were comparable with PDMS and TCPS, respectively. Moreover, using primary rat hepatocytes, we demonstrated maintained high levels of liver function at least for 1 week on PMP, with its low chemical sorption and high oxygen permeability being key factors in both revealing the potential of primary cultured hepatocytes and in performing an accurate evaluation of hepatic metabolisms. Taken together, we conclude that PMP is a superior alternative to both PDMS and TCPS, and a promising material for a variety of drug testing systems.

A parallelized, perfused 3D triculture model of leukemia for in vitro drug testing of chemotherapeutics

Leukemia patients undergo chemotherapy to combat the leukemic cells (LCs) in the bone marrow. During therapy not only the LCs, but also the blood-producing hematopoietic stem and progenitor cells (HSPCs) may be destroyed. Chemotherapeutics targeting only the LCs are urgently needed to overcome this problem and minimize life-threatening side-effects. Predictive in vitro drug testing systems allowing simultaneous comparison of various experimental settings would enhance the efficiency of drug development. Here, we present a three-dimensional (3D) human leukemic bone marrow model perfused using a magnetic, parallelized culture system to ensure media exchange. Chemotherapeutic treatment of the acute myeloid leukemia cell line KG-1a in 3D magnetic hydrogels seeded with mesenchymal stem/stromal cells (MSCs) revealed a greater resistance of KG-1a compared to 2D culture. In 3D tricultures with HSPCs, MSCs and KG-1a, imitating leukemic bone marrow, HSPC proliferation decreased while KG-1a cells remained unaffected post treatment. Non-invasive metabolic profiling enabled continuous monitoring of the system. Our results highlight the importance of using biomimetic 3D platforms with proper media exchange and co-cultures for creating in vivo-like conditions to enable in vitro drug testing. This system is a step towards drug testing in biomimetic, parallelized in vitro approaches, facilitating the discovery of new anti-leukemic drugs.

Human Mesenchymal Stem Cell-Derived Miniature Joint System for Disease Modeling and Drug Testing.

Diseases of the knee joint such as osteoarthritis (OA) affect all joint elements. An in vitro human cell‐derived microphysiological system capable of simulating intraarticular tissue crosstalk is desirable for studying etiologies/pathogenesis of joint diseases and testing potential therapeutics. Herein, a human mesenchymal stem cell‐derived miniature joint system (miniJoint) is generated, in which engineered osteochondral complex, synovial‐like fibrous tissue, and adipose tissue are integrated into a microfluidics‐enabled bioreactor. This novel design facilitates different tissues communicating while still maintaining their respective phenotypes. The miniJoint exhibits physiologically relevant changes when exposed to interleukin‐1β mediated inflammation, which are similar to observations in joint diseases in humans. The potential of the miniJoint in predicting in vivo efficacy of drug treatment is confirmed by testing the “therapeutic effect” of the nonsteroidal anti‐inflammatory drug, naproxen, as well as four other potential disease‐modifying OA drugs. The data demonstrate that the miniJoint recapitulates complex tissue interactions, thus providing a robust organ chip model for the study of joint pathology and the development of novel therapeutic interventions.

Central nervous system organoids for modeling neurodegenerative diseases.

Recent advances in induced pluripotent stem cell (iPSC) technology have allowed researchers to generate neurodegenerative disease-specific iPSCs and use the cells to derive a variety of relevant cell populations for laboratory modeling and drug testing. Nevertheless, these efforts have faced challenges related to immaturity and lack of complex developmental niches in the derived cell populations, limiting the utility of these in vitro models of neurodegenerative disease. Such limitations may be overcome by using human iPSC technology to generate three-dimensional (3D) brain organoids, which better recapitulate in vivo tissue architecture than traditional neuronal cultures to provide more complex and representative disease models and drug testing systems. In this review, we focus on the application of pluripotent stem cell-derived central nervous system (CNS) organoids to model neurodegenerative diseases. We first summarize recent progress in generating and characterizing various CNS organoids from pluripotent stem cells. We then review the application of CNS organoids for modeling several different human neurodegenerative diseases. We also describe several novel pathological mechanisms and drugs that were studied using patient iPSC-derived CNS organoids. Finally, we discuss remaining challenges and emerging opportunities for the use of 3D brain organoids for in vitro modeling of CNS development and neurodegeneration.

From organ-on-chip to body-on-chip: The next generation of microfluidics platforms for in vitro drug efficacy and toxicity testing.

The high failure rate in drug development is often attributed to the lack of accurate pre-clinical models that may lead to false discoveries and inconclusive data when the compounds are eventually tested in clinical phase. With the evolution of cell culture technologies, drug testing systems have widely improved, and today, with the emergence of microfluidics devices, drug screening seems to be at the dawn of an important revolution. An organ-on-chip allows the culture of living cells in continuously perfused microchambers to reproduce physiological functions of a particular tissue or organ. The advantages of such systems are not only their ability to recapitulate the complex biochemical interactions between different human cell types but also to incorporate physical forces, including shear stress and mechanical stretching or compression. To improve this model, and to reproduce the absorption, distribution, metabolism, and elimination process of an exogenous compound, organ-on-chips can even be linked fluidically to mimic physiological interactions between different organs, leading to the development of body-on-chips. Although these technologies are still at a young age and need to address a certain number of limitations, they already demonstrated their relevance to study the effect of drugs or toxins on organs, displaying a similar response to what is observed in vivo. The purpose of this review is to present the evolution from organ-on-chip to body-on-chip, examine their current use for drug testing and discuss their advantages and future challenges they will face in order to become an essential pillar of pharmaceutical research.

Dealing with Ethical Issues in Clinical Trials: The ussr in the Global Context

Abstract This paper discusses several ethical issues related to clinical trials within the Soviet system of drug development and testing, which reflected larger ideological principles of healthcare organization in the ussr, with its focus on eradicating market elements from drug development. The centralized state-controlled system was thought to combat such drawbacks of free-market drug development as high prices and aggressive advertising; also to discourage the duplication of research by numerous independent actors that was perceived to be common in capitalist countries. Another significant ethical issue was the Soviet emphasis on the unity of scientific research and clinical treatment. Their strict separation, introduced to support normative standards defined by the U.S. pharmaceutical drug testing system, was rejected in the ussr where knowledge of new treatment options came from treatment practice, not laboratory-like experimental conditions of randomized controlled double-blind trials. The Soviet design was closer to so-called ‘pragmatic trials’ that focus on solving ‘real-life’ problems in clinical practice. Not all ethical problems were successfully addressed in the Soviet model, where there were always significant gaps between neatly postulated theory and messy clinical practice. The unity of scientific research and clinical practice was difficult to achieve. Archival research shows potential ethical issues related to geographic disparities in carrying out clinical trials, and the importance of personal and informal connections in the Soviet model.

The Ultrastructural Analysis of Human Colorectal Cancer Stem Cell-Derived Spheroids and Their Mouse Xenograft Shows That the Same Cells Types Have Different Ratios.

Simple SummaryA comparative ultrastructural and flow cytometric analysis of colorectal cancer-derived spheroids and their mouse xenografts showed that they both contain the same cell types but with different ratios, reflecting the interaction of cancer cells, respectively, with the in vitro and in vivo microenvironment.Spheroids from primary colorectal cancer cells and their mice xenografts have emerged as useful preclinical models for cancer research as they replicate tumor features more faithfully as compared to cell lines. While 3D models provide a reliable system for drug discovery and testing, their structural complexity represents a challenge and their structure-function relationships are only partly understood. Here, we present a comparative ultrastructural and flow citometric analysis of patient colorectal cancer-derived spheroids and their mice xenografts. Ultrastructural observations highlighted that multicellular spheroids and their xenografts contain the same cancer cell types but with different ratios, specifically multicellular spheroids were enriched in cells with a stem-like phenotype, while xenografts had an increased amount of lipid droplets-containing cells. The flow cytometric analysis for stem cell marker and activity showed enrichment of stem-like cells presence and activity in spheroids while xenografts had the inverse response. Our results evidence the effects on cancer cells of different in vitro and in vivo microenvironments. Those differences have to be paid into account in designing innovative experimental models for personalized drug testing.

Neuroblastoma and DIPG Organoid Coculture System for Personalized Assessment of Novel Anticancer Immunotherapies.

Cancer immunotherapy has transformed the landscape of adult cancer treatment and holds a great promise to treat paediatric malignancies. However, in vitro test coculture systems to evaluate the efficacy of immunotherapies on representative paediatric tumour models are lacking. Here, we describe a detailed procedure for the establishment of an ex vivo test coculture system of paediatric tumour organoids and immune cells that enables assessment of different immunotherapy approaches in paediatric tumour organoids. We provide a step-by-step protocol for an efficient generation of patient-derived diffuse intrinsic pontine glioma (DIPG) and neuroblastoma organoids stably expressing eGFP-ffLuc transgenes using defined serum-free medium. In contrast to the chromium-release assay, the new platform allows for visualization, monitoring and robust quantification of tumour organoid cell cytotoxicity using a non-radioactive assay in real-time. To evaluate the utility of this system for drug testing in the paediatric immuno-oncology field, we tested our in vitro assay using a clinically used immunotherapy strategy for children with high-risk neuroblastoma, dinutuximab (anti-GD2 monoclonal antibody), on GD2 proficient and deficient patient-derived neuroblastoma organoids. We demonstrated the feasibility and sensitivity of our ex vivo coculture system using human immune cells and paediatric tumour organoids as ex vivo tumour models. Our study provides a novel platform for personalized testing of potential anticancer immunotherapies for aggressive paediatric cancers such as neuroblastoma and DIPG.