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The COVID-19 pandemic has presented an unprecedented challenge to the healthcare system. Identifying the genomics and clinical biomarkers for effective patient stratification and management is critical to controlling the spread of the disease. Omics datasets provide a wealth of information that can aid in understanding the underlying molecular mechanisms of COVID-19 and identifying potential biomarkers for patient stratification. Artificial intelligence (AI) and machine learning (ML) algorithms have been increasingly used to analyze large-scale omics and clinical datasets for patient stratification. In this manuscript, we demonstrate the recent advances and predictive accuracies in AI- and ML-based patient stratification modeling linking omics and clinical biomarker datasets, focusing on COVID-19 patients. Our ML model not only demonstrates that clinical features are enough of an indicator of COVID-19 severity and survival, but also infers what clinical features are more impactful, which makes our approach a useful guide for clinicians for prioritization best-fit therapeutics for a given cohort of patients. Moreover, with weighted gene network analysis, we are able to provide insights into gene networks that have a significant association with COVID-19 severity and clinical features. Finally, we have demonstrated the importance of clinical biomarkers in identifying high-risk patients and predicting disease progression.
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Inteligencia Artificial , COVID-19 , Humanos , COVID-19/genética , Medicina de Precisión , Pandemias , Aprendizaje Automático , BiomarcadoresRESUMEN
Though the portfolio of medicines that are extending and improving the lives of patients continues to grow, drug discovery and development remains a challenging business on its best day. Safety liabilities are a significant contributor to development attrition where the costliest liabilities to both drug developers and patients emerge in late development or post-marketing. Animal studies are an important and influential contributor to the current drug discovery and development paradigm intending to provide evidence that a novel drug candidate can be used safely and effectively in human volunteers and patients. However, translational gaps-such as toxicity in patients not predicted by animal studies-have prompted efforts to improve their effectiveness, especially in safety assessment. More holistic monitoring and "digitalization" of animal studies has the potential to enrich study outcomes leading to datasets that are more computationally accessible, translationally relevant, replicable, and technically efficient. Continuous monitoring of animal behavior and physiology enables longitudinal assessment of drug effects, detection of effects during the animal's sleep and wake cycles and the opportunity to detect health or welfare events earlier. Automated measures can also mitigate human biases and reduce subjectivity. Reinventing a conservative, standardized, and traditional paradigm like drug safety assessment requires the collaboration and contributions of a broad and multi-disciplinary stakeholder group. In this perspective, we review the current state of the field and discuss opportunities to improve current approaches by more fully leveraging the power of sensor technologies, artificial intelligence (AI), and animal behavior in a home cage environment.
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Historical data from control groups in animal toxicity studies is currently mainly used for comparative purposes to assess validity and robustness of study results. Due to the highly controlled environment in which the studies are performed and the homogeneity of the animal collectives it has been proposed to use the historical data for building so-called virtual control groups, which could replace partly or entirely the concurrent control. This would constitute a substantial contribution to the reduction of animal use in safety studies. Before the concept can be implemented, the prerequisites regarding data collection, curation and statistical evaluation together with a validation strategy need to be identified to avoid any impairment of the study outcome and subsequent consequences for human risk assessment. To further assess and develop the concept of virtual control groups the transatlantic think tank for toxicology (t4) sponsored a workshop with stakeholders from the pharmaceutical and chemical industry, academia, FDA, pharmaceutical, contract research organizations (CROs), and non-governmental organizations in Washington, which took place in March 2023. This report summarizes the current efforts of a European initiative to share, collect and curate animal control data in a centralized database and the first approaches to identify optimal matching criteria between virtual controls and the treatment arms of a study as well as first reflections about strategies for a qualification procedure and potential pitfalls of the concept.
Animal safety studies are usually performed with three groups of animals where increasing amounts of the test chemical are given to the animals and one control group where the animals do not receive the test chemical. The design of such studies, the characteristics of the animals, and the measured parameters are often very similar from study to study. Therefore, it has been suggested that measurement data from the control groups could be reused from study to study to lower the total number of animals per study. This could reduce animal use by up to 25% for such standardized studies. A workshop was held to discuss the pros and cons of such a concept and what would have to be done to implement it without threatening the reliability of the study outcome or the resulting human risk assessment.
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Investigación , Animales , Grupos Control , Preparaciones FarmacéuticasRESUMEN
Complex in vitro models (CIVM) offer the potential to improve pharmaceutical clinical drug attrition due to safety and/ or efficacy concerns. For this technology to have an impact, the establishment of robust characterization and qualification plans constructed around specific contexts of use (COU) is required. This article covers the output from a workshop between the Food and Drug Administration (FDA) and Innovation and Quality Microphysiological Systems (IQ MPS) Affiliate. The intent of the workshop was to understand how CIVM technologies are currently being applied by pharmaceutical companies during drug development and are being tested at the FDA through various case studies in order to identify hurdles (real or perceived) to the adoption of microphysiological systems (MPS) technologies, and to address evaluation/qualification pathways for these technologies. Output from the workshop includes the alignment on a working definition of MPS, a detailed description of the eleven CIVM case studies presented at the workshop, in-depth analysis, and key take aways from breakout sessions on ADME (absorption, distribution, metabolism, and excretion), pharmacology, and safety that covered topics such as qualification and performance criteria, species differences and concordance, and how industry can overcome barriers to regulatory submission of CIVM data. In conclusion, IQ MPS Affiliate and FDA scientists were able to build a general consensus on the need for animal CIVMs for preclinical species to better determine species concordance. Furthermore, there was acceptance that CIVM technologies for use in ADME, pharmacology and safety assessment will require qualification, which will vary depending on the specific COU.
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Alternativas a las Pruebas en Animales , Dispositivos Laboratorio en un Chip , Animales , Evaluación Preclínica de Medicamentos , Industria Farmacéutica , Preparaciones Farmacéuticas/metabolismo , Estados Unidos , United States Food and Drug AdministrationRESUMEN
To understand the growing needs of an aging human population, there is demand for scalable and reproducible approaches to study animal models of aging and to test novel therapeutic interventions. We investigated the sensitivity and utility of a continuous monitoring platform and its digital biomarkers (motion, breathing rate, and wheel running) to evaluate behavioral and physiological differences between "young" (12 weeks) and "old" (23 months) male C57BL/6J mice with or without running wheels in the home cage. Compared to young mice, old mice showed marked reductions in motion and breathing rate, as well as altered circadian rhythms. Mice without running wheels possessed lower breathing rates compared to their counterparts with running wheels. Digital biomarkers showed age-dependent changes in response to routine procedures (cage changes and blood sampling) and alterations in subjects that unexpectedly reached endpoint. Continuous collection of digital biomarkers in the home cage can enhance current approaches by providing unbiased longitudinal monitoring for large-scale aging studies.
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Envejecimiento/fisiología , Conducta Animal/fisiología , Biomarcadores/análisis , Monitoreo Fisiológico/instrumentación , Actividad Motora/fisiología , Animales , Automatización , Ritmo Circadiano/fisiología , Determinación de Punto Final , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Animales , RespiraciónRESUMEN
In drug discovery and development, traditional assessment of human patients and preclinical subjects occurs at limited time points in potentially stressful surroundings (i.e., the clinic or a test arena), which can impact data quality and welfare. However, recent advances in remote digital monitoring technologies enable the assessment of human patients and preclinical subjects across multiple time points in familiar surroundings. The ability to monitor a patient throughout disease progression provides an opportunity for more relevant and efficient diagnosis as well as improved assessment of drug efficacy and safety. In preclinical in vivo animal models, these digital technologies allow for continuous, longitudinal, and non-invasive monitoring in the home environment. This manuscript provides an overview of digital monitoring technologies for use in preclinical studies including their history and evolution, current engagement through use cases, and impact of digital biomarkers (DBs) on drug discovery and the 3Rs. We also discuss barriers to implementation and strategies to overcome them. Finally, we address data consistency and technology standards from the perspective of technology providers, end-users, and subject matter experts. Overall, this review establishes an improved understanding of the value and implementation of digital biomarker (DB) technologies in preclinical research.
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Cord blood transplantation (CBT) with units containing total nucleated cell (TNC) dose >2.5 x 10(7)/kg is associated with improved engraftment and decreased transplant-related mortality. For many adults no single cord blood units are available that meet the cell dose requirements. We developed a dog model of CBT to evaluate approaches to overcome the problem of low cell dose cord blood units. This study primarily compared double- versus single-unit CBT. Unrelated dogs were bred and cord blood units were harvested. We identified unrelated recipients that were dog leukocyte antigen (DLA)-88 (class I) and DLA-DRB1 (class II) allele-matched with cryopreserved units. Each unit contained Asunto(s)
Trasplante de Células Madre de Sangre del Cordón Umbilical/métodos
, Animales
, Trasplante de Células Madre de Sangre del Cordón Umbilical/efectos adversos
, Modelos Animales de Enfermedad
, Perros
, Femenino
, Rechazo de Injerto/inmunología
, Masculino
, Tasa de Supervivencia
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Safety related drug failures continue to be a challenge for pharmaceutical companies despite the numerous complex and lengthy in vitro assays and in vivo studies that make up the typical safety screening funnel. A lack of complete translation of animal data to humans can explain some of those shortcomings. Differences in sensitivity and drug disposition between animals and humans may also play a role. Many gaps exist for potential target tissues of drugs that cannot be adequately modeled in vitro. Microphysiological systems (MPS) may help to better model these target tissues and provide an opportunity to better assess some aspects of human safety prior to clinical studies. There is hope that these systems can supplement current preclinical drug safety and disposition evaluations, filling gaps and enhancing our ability to predict and understand human relevant toxicities. The International Consortium for Innovation and Quality in Pharmaceutical Development (IQ) MPS Affiliate is a group of pharmaceutical industry scientists who seek to expedite appropriate characterization and incorporation of MPS to potentially improve drug safety assessment and provide safer and more effective medicines to patients. In keeping with this mission, the IQ MPS Affiliate scientists have prepared a series of organotypic manuscripts for several key drug safety and disposition target tissues (lung, liver, kidney, skin, gastrointestinal, cardiovascular, and blood brain barrier/central nervous system). The goal of these manuscripts is to provide key information related to likely initial contexts of use (CoU) and key characterization data needed for incorporation of MPS in pharmaceutical safety screening including a list of characteristic functions, cell types, toxicities, and test agents (representing major mechanisms of toxicity) that can be used by MPS developers. Additional manuscripts focusing on testing biologically based therapeutics and ADME considerations have been prepared as part of this effort. These manuscripts focus on general needs for assessing biologics and ADME endpoints and include similar information to the tissue specific manuscripts where appropriate. The current manuscript is an introduction to several general concepts related to pharmaceutical industry needs with regard to MPS application and other MPS concepts that apply across the organ specific manuscripts.
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Hígado , Preparaciones Farmacéuticas , Animales , Barrera Hematoencefálica , Humanos , Riñón , PulmónRESUMEN
Assessments of respiratory response and animal activity are useful endpoints in drug pharmacology and safety research. We investigated whether continuous, direct monitoring of breathing rate and body motion in animals in the home cage using the Vum Digital Smart House can complement standard measurements in enabling more granular detection of the onset and severity of physiologic events related to lung injury in a well-established rodent model of paraquat (PQ) toxicity. In rats administered PQ, breathing rate was significantly elevated while body motion was significantly reduced following dosing and extending throughout the 14-day study duration for breathing rate and at least 5 days for both nighttime and daytime body motion. Time course differences in these endpoints in response to the potential ameliorative test article bardoxolone were also readily detected. More complete than standard in-life measurements, breathing rate and body motion tracked injury progression continuously over the full study time period and aligned with, and informed on interval changes in clinical pathology. In addition, breathing rates correlated with terminal pathology measurements, such as normalized lung weights and histologic alveolar damage and edema. This study is a preliminary evaluation of the technology; our results demonstrate that continuously measured breathing rate and body motion served as physiologically relevant readouts to assess lung injury progression and drug response in a respiratory injury animal model.
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The animal research community faces a shortage of surgical training opportunities along with an increasing demand for expertise in surgical techniques. One possible means of overcoming this challenge is the use of computer-based or electronic learning (e-learning) to disseminate material to a broad range of animal users. E-learning platforms can take many different forms, ranging from simple text documents that are posted online to complex virtual courses that incorporate dynamic video or audio content and in which students and instructors can interact in real time. The authors present an overview of e-learning and discuss its potential benefits as a supplement to hands-on rodent surgical training. They also discuss a few basic considerations in developing and implementing electronic courses.
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Animales de Laboratorio/cirugía , Instrucción por Computador/métodos , Ciencia de los Animales de Laboratorio/educación , Cirugía Veterinaria/educación , Animales , Curriculum , HumanosRESUMEN
The animal research community comprises members from a wide variety of backgrounds, some of whom must learn basic surgical skills. Though demand for animal research personnel who have surgical skills is increasing, surgical training opportunities are becoming more scarce. Electronic learning or e-learning platforms can be used as an adjunct to hands-on surgical training. Course developers can adapt these e-learning courses to fit the needs of participants who have varying levels of expertise. The authors outline the steps involved in developing an effective e-learning surgical course. They also describe how to use various equipment and software products to help implement e-learning courses. Though the authors focus on developing surgical courses, course developers could apply the general steps outlined by the authors when developing any e-learning course.
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Animales de Laboratorio/cirugía , Instrucción por Computador/métodos , Cirugía General/educación , Animales , HumanosRESUMEN
Studies using nonhuman primate embryonic stem (ES) cells will facilitate the translation of basic ES cell research to clinical use by providing a large animal model for in vivo testing. Unfortunately, nonhuman primate ES cells do not survive well following cryopreservation, a problem that limits the quantity and quality of these cells for research. More lines have to be established to fulfill demand, and thawed aliquots must go through more passages to generate adequate numbers. In addition, suboptimal cryopreservation can induce epigenetic changes and impose a selection bias for their outgrowth. Therefore, defining the optimal cryopreservation technique for nonhuman primate ES cells is critical for the further development of this research. To address this problem, we tested various cryoprotectants as well as cryopreservation procedures in an attempt to define a protocol that yields high viability with retention of ES cell phenotype and function. Here, we report a freezing protocol that preserves the intercellular attachments that are vital to primate ES cell function. We describe a slow, controlled-rate cooling protocol with ice crystal induction that increased the survival rate of ES cells from <22% to >90%. Preserved cells retained a normal karyotype and did not lose their ability to express markers of undifferentiated ES cells.
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Criopreservación/métodos , Crioprotectores/metabolismo , Células Madre Embrionarias/fisiología , Animales , Biomarcadores/metabolismo , Línea Celular , Forma de la Célula , Supervivencia Celular , Células Madre Embrionarias/citología , Humanos , Cariotipificación , Macaca mulatta , MasculinoRESUMEN
Freeze storage of human embryonic stem (hES) cells has not proven effective using the methods employed for mouse ES (mES) cells, while rhesus ES (rhES) cells are only modestly effectively frozen using common mES cell methods. Because human and rhES cells are passaged and frozen in clusters that approximate the size of embryos, we employed a mammalian embryo freezing method to cryopreserve primate ES cells. This protocol involves freezing in a dimethylsulfoxide cryoprotectant using straws. An ice crystal seed is induced at -10 degrees C followed by controlled cooling at -1 degrees C per minute down to -33 degrees C with a plunge from there directly into liquid nitrogen (LN2) at -196 degrees C. Thaw is effected rapidly by moving the frozen cells directly from LN2 into a water bath and placing directly into culture medium without step-wise cryoprotectant removal. Using this protocol, we have increased the survival of human ES cells from < or = 1 to approximately 80% and rhES cells from approximately 30 to > or = 90%. Thus, this protocol describes a technically simple but effective means of long-term storage of primate ES cells.
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Técnicas de Cultivo de Célula/métodos , Criopreservación/métodos , Células Madre Embrionarias/citología , Animales , Células Cultivadas , Dimetilsulfóxido , Congelación , Humanos , Ratones , PrimatesRESUMEN
Personnel working with laboratory animals are required by laws and guidelines to be trained and qualified to perform biomethodologic procedures. The assessment of competency and proficiency is a vital component of a laboratory animal training program, because this process confirms that the trainees have met the learning objectives for a particular procedure. The approach toward qualification assessment differs between organizations because laws and guidelines do not outline how the assessment should be performed or which methods and tools should be used. Assessment of clinical and surgical medicine has received considerable attention over the last few decades and has progressed from simple subjective methods to well-defined and objective methods of assessing competency. Although biomethodology competency and proficiency assessment is discussed in the literature, a standard and objective assessment method has not yet been developed. The development and implementation of an objective and standardized biomethodologic assessment program can serve as a tool to improve standards, ensure consistent training, and decrease research variables yet ensure animal welfare. Here we review the definition and goals of training and assessment, review assessment methods, and propose a method to develop a standard and objective assessment program for the laboratory animal science field, particularly training departments and IACUC.
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Ciencia de los Animales de Laboratorio/educación , Comités de Atención Animal , Bienestar del Animal , Animales , Animales de Laboratorio , Ciencia de los Animales de Laboratorio/normasRESUMEN
The refinement of surgical techniques represents a key opportunity to improve the welfare of laboratory rodents, while meeting legal and ethical obligations. Current methods used for monitoring intra-abdominal disease progression in rodents usually involve euthanasia at various time-points for end of study, one-time individual tissue collections. Most rodent organ tumour models are developed by the introduction of tumour cells via laparotomy or via ultrasound-guided indirect visualization. Ischaemic rodent models are often generated using laparotomies. This approach requires a high number of rodents, and in some instances introduces high degrees of morbidity and mortality, thereby increasing study variability and expense. Most importantly, most laparotomies do not promote the highest level of rodent welfare. Recent improvements in laparoscopic equipment and techniques have enabled the adaptation of laparoscopy for rodent procedures. Laparoscopy, which is considered the gold standard for many human abdominal procedures, allows for serial biopsy collections from the same animal, results in decreased pain and tissue trauma as well as quicker postsurgical recovery, and preserves immune function in comparison to the same procedures performed by laparotomy. Laparoscopy improves rodent welfare, decreases inter-animal variability, thereby reducing the number of required animals, allows for the replacement of larger species, decreases expense and improves data yield. This review article compares rodent laparotomy and laparoscopic surgical methods, and describes the utilization of laparoscopy for the development of cancer models and assessment of disease progression to improve data collection and animal welfare. In addition, currently available rodent laparoscopic equipment and instrumentation are presented.
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Bienestar del Animal , Animales de Laboratorio , Laparoscopía/instrumentación , Laparoscopía/métodos , Laparotomía/métodos , Monitoreo Fisiológico/métodos , Roedores , AnimalesRESUMEN
The development of new rodent models of human disease and advances in surgical equipment and technologies have increased the demand for expertise in rodent surgery. Because of the limited availability of rodent surgical training courses, electronic (e-) learning is presented as an alternative to in-person education and as a means to hone the expertise of current surgeons in biomedical research, similar to e-learning applications for human surgery training. Translating this model to the biomedical research field provides participants with an opportunity to train themselves on rodent surgical techniques prior to operating on live models. An e-learning rodent surgery course was incorporated into a training class of undergraduate (n = 39) and graduate (n = 12) laboratory animal students, and a portion of the course was presented to laboratory animal professionals (n = 15). The effectiveness of the method was evaluated using written examination and postcourse surveys. The exam data demonstrated that the e-learning course transferred knowledge comparable to a lecture course on surgery that was presented in-person. Students responded favorably to videos, step-by-step photographs of surgical procedures, and the ready accessibility of the course. Critiques included the need to improve video resolution and quality of the voice-overs. These results support the continued development and implementation of electronic rodent surgical technique courses for use in laboratory animal and biomedical research communities.
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Instrucción por Computador/métodos , Cirugía General/educación , Multimedia , Roedores/cirugía , Animales , InternetRESUMEN
Although hematopoietic cell transplantation (HCT) is generally accomplished using a single donor, multiple donors have been used to enhance the speed of engraftment, particularly in the case of umbilical cord blood grafts. Here we posed the question in the canine HCT model whether stable dual-donor chimerism could be established using 2 DLA-identical donors. We identified 8 DLA-identical littermate triplets in which the marrow recipients received 2 Gy total body irradiation followed by marrow infusions from 2 donors and postgrafting immunosuppression. All 8 dogs showed initial "trichimerism," which was sustained in 5 dogs, while 2 dogs rejected one of the allografts and remained mixed chimeras, and 1 dog rejected both allografts. Immune function in one trichimeric dog, as tested by mixed leukocyte culture response and antibody response to sheep red blood cells, was found to be normal. Five dogs received kidney grafts from one of their respective marrow donors at least 6 months after HCT without immunosuppressive drugs, and grafts in 4 dogs are surviving without rejection. In summary, following nonmyeloablative conditioning, simultaneous administration of marrow grafts from 2 DLA-identical littermates could result in sustained trichimerism, and immunologic tolerance could include a kidney graft from one of the marrow donors.