المؤلفون: McKinley, Jonathan P, O'Connell, Grace D

مصطلحات موضوعية: Engineering, Biomedical Engineering, Pain Research, Bioengineering, Chronic Pain, Biotechnology, Animals, Humans, Intervertebral Disc Degeneration, Quality of Life, Organ Culture Techniques, Intervertebral Disc, Bioreactors, Bioreactor, Complex loading, Disc degeneration, Intervertebral disc, Mechanical Engineering, Human Movement and Sports Sciences, Biomedical engineering, Sports science and exercise

الوصف: Lower back pain continues to be a global epidemic, limiting quality of life and ability to work, due in large part to symptomatic disc degeneration. Development of more effective and less invasive biological strategies are needed to treat disc degeneration. In vitro models such as macro- or micro-bioreactors or mechanically active organ-chips hold great promise in reducing the need for animal studies that may have limited clinical translatability, due to harsher and more complex mechanical loading environments in human discs than in most animal models. This review highlights the complex loading conditions of the disc in situ, evaluates state-of-the-art designs for applying such complex loads across multiple length scales, from macro-bioreactors that load whole discs to organ-chips that aim to replicate cellular or engineered tissue loading. Emphasis was placed on the rapidly evolving more customizable organ-chips, given their greater potential for studying the progression and treatment of symptomatic disc degeneration. Lastly, this review identifies new trends and challenges for using organ-chips to assess therapeutic strategies.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/0s8150gpTest

المؤلفون: Karzbrun, Eyal, Khankhel, Aimal H, Megale, Heitor C, Glasauer, Stella MK, Wyle, Yofiel, Britton, George, Warmflash, Aryeh, Kosik, Kenneth S, Siggia, Eric D, Shraiman, Boris I, Streichan, Sebastian J

المصدر: Nature. 599(7884)

مصطلحات موضوعية: Biological Sciences, Engineering, Biomedical Engineering, Regenerative Medicine, Stem Cell Research - Embryonic - Human, Stem Cell Research, Pediatric, Stem Cell Research - Nonembryonic - Human, Neurosciences, Biotechnology, Underpinning research, 1.1 Normal biological development and functioning, Generic health relevance, Ectoderm, Humans, Models, Biological, Morphogenesis, Neural Plate, Neural Tube, Neural Tube Defects, Organ Culture Techniques, Regeneration, Stem Cells, General Science & Technology

الوصف: Understanding human organ formation is a scientific challenge with far-reaching medical implications1,2. Three-dimensional stem-cell cultures have provided insights into human cell differentiation3,4. However, current approaches use scaffold-free stem-cell aggregates, which develop non-reproducible tissue shapes and variable cell-fate patterns. This limits their capacity to recapitulate organ formation. Here we present a chip-based culture system that enables self-organization of micropatterned stem cells into precise three-dimensional cell-fate patterns and organ shapes. We use this system to recreate neural tube folding from human stem cells in a dish. Upon neural induction5,6, neural ectoderm folds into a millimetre-long neural tube covered with non-neural ectoderm. Folding occurs at 90% fidelity, and anatomically resembles the developing human neural tube. We find that neural and non-neural ectoderm are necessary and sufficient for folding morphogenesis. We identify two mechanisms drive folding: (1) apical contraction of neural ectoderm, and (2) basal adhesion mediated via extracellular matrix synthesis by non-neural ectoderm. Targeting these two mechanisms using drugs leads to morphological defects similar to neural tube defects. Finally, we show that neural tissue width determines neural tube shape, suggesting that morphology along the anterior-posterior axis depends on neural ectoderm geometry in addition to molecular gradients7. Our approach provides a new route to the study of human organ morphogenesis in health and disease.

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الوصول الحر: https://escholarship.org/uc/item/8fg4s73wTest

المؤلفون: Berg, Elizabeth L, Jami, Shekib A, Petkova, Stela P, Berz, Annuska, Fenton, Timothy A, Lerch, Jason P, Segal, David J, Gray, John A, Ellegood, Jacob, Wöhr, Markus, Silverman, Jill L

المصدر: Journal of Neuroscience. 41(42)

مصطلحات موضوعية: Biomedical and Clinical Sciences, Neurosciences, Orphan Drug, Pediatric, Intellectual and Developmental Disabilities (IDD), Rare Diseases, Brain Disorders, Mental Health, Aetiology, 2.1 Biological and endogenous factors, Neurological, Mental health, Angelman Syndrome, Animals, Brain, Disease Models, Animal, Female, Gene Deletion, Laughter, Male, Microcephaly, Organ Culture Techniques, Protein Biosynthesis, Rats, Rats, Sprague-Dawley, Rats, Transgenic, Reflex, Startle, Social Behavior, Ubiquitin-Protein Ligases, Vocalization, Animal, Angelman syndrome, behavior, play, rat, Ube3a, ultrasonic vocalization, Medical and Health Sciences, Psychology and Cognitive Sciences, Neurology & Neurosurgery

الوصف: Angelman syndrome (AS) is a rare genetic neurodevelopmental disorder characterized by intellectual disabilities, motor and balance deficits, impaired communication, and a happy, excitable demeanor with frequent laughter. We sought to elucidate a preclinical outcome measure in male and female rats that addressed communication abnormalities of AS and other neurodevelopmental disorders in which communication is atypical and/or lack of speech is a core feature. We discovered, and herein report for the first time, excessive laughter-like 50 kHz ultrasonic emissions in the Ube3amat-/pat+ rat model of AS, which suggests an excitable, playful demeanor and elevated positive affect, similar to the demeanor of individuals with AS. Also in line with the AS phenotype, Ube3amat-/pat+ rats demonstrated aberrant social interactions with a novel partner, distinctive gait abnormalities, impaired cognition, an underlying LTP deficit, and profound reductions in brain volume. These unique, robust phenotypes provide advantages compared with currently available mouse models and will be highly valuable as outcome measures in the evaluation of therapies for AS.SIGNIFICANCE STATEMENT Angelman syndrome (AS) is a severe neurogenetic disorder for which there is no cure, despite decades of research using mouse models. This study used a recently developed rat model of AS to delineate disease-relevant outcome measures to facilitate therapeutic development. We found the rat to be a strong model of AS, offering several advantages over mouse models by exhibiting numerous AS-relevant phenotypes, including overabundant laughter-like vocalizations, reduced hippocampal LTP, and volumetric anomalies across the brain. These findings are unconfounded by detrimental motor abilities and background strain, issues plaguing mouse models. This rat model represents an important advancement in the field of AS, and the outcome metrics reported herein will be central to the therapeutic pipeline.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/8fz2b3z9Test

المؤلفون: Jami, Shekib A, Cameron, Scott, Wong, Jonathan M, Daly, Emily R, McAllister, A Kimberley, Gray, John A

المصدر: Journal of Neurophysiology. 126(1)

مصطلحات موضوعية: Biomedical and Clinical Sciences, Biological Psychology, Neurosciences, Psychology, Pharmacology and Pharmaceutical Sciences, Brain Disorders, Pediatric, Schizophrenia, Mental Health, 2.1 Biological and endogenous factors, Aetiology, Mental health, Neurological, Animals, Excitatory Postsynaptic Potentials, Female, GABAergic Neurons, Hippocampus, Inhibitory Postsynaptic Potentials, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Organ Culture Techniques, Racemases and Epimerases, Receptors, N-Methyl-D-Aspartate, Synapses, E, I balance, GABA, inhibition, NMDA receptor, SRR, E/I balance, Medical and Health Sciences, Psychology and Cognitive Sciences, Neurology & Neurosurgery, Biomedical and clinical sciences, Health sciences

الوصف: There is substantial evidence that both N-methyl-D-aspartate receptor (NMDAR) hypofunction and dysfunction of GABAergic neurotransmission contribute to schizophrenia, though the relationship between these pathophysiological processes remains largely unknown. Although models using cell-type-specific genetic deletion of NMDARs have been informative, they display overly pronounced phenotypes extending beyond those of schizophrenia. Here, we used the serine racemase knockout (SRKO) mice, a model of reduced NMDAR activity rather than complete receptor elimination, to examine the link between NMDAR hypofunction and decreased GABAergic inhibition. The SRKO mice, in which there is a >90% reduction in the NMDAR coagonist d-serine, exhibit many of the neurochemical and behavioral abnormalities observed in schizophrenia. We found a significant reduction in inhibitory synapses onto CA1 pyramidal neurons in the SRKO mice. This reduction increases the excitation/inhibition balance resulting in enhanced synaptically driven neuronal excitability without changes in intrinsic excitability. Consistently, significant reductions in inhibitory synapse density in CA1 were observed by immunohistochemistry. We further show, using a single-neuron genetic deletion approach, that the loss of GABAergic synapses onto pyramidal neurons observed in the SRKO mice is driven in a cell-autonomous manner following the deletion of SR in individual CA1 pyramidal cells. These results support a model whereby NMDAR hypofunction in pyramidal cells disrupts GABAergic synapses leading to disrupted feedback inhibition and impaired neuronal synchrony.NEW & NOTEWORTHY Recently, disruption of excitation/inhibition (E/I) balance has become an area of considerable interest for psychiatric research. Here, we report a reduction in inhibition in the serine racemase knockout mouse model of schizophrenia that increases E/I balance and enhances synaptically driven neuronal excitability. This reduced inhibition was driven cell-autonomously in pyramidal cells lacking serine racemase, suggesting a novel mechanism for how chronic NMDA receptor hypofunction can disrupt information processing in schizophrenia.

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الوصول الحر: https://escholarship.org/uc/item/125438ssTest

المؤلفون: Oyler-Yaniv, Jennifer, Oyler-Yaniv, Alon, Maltz, Evan, Wollman, Roy

المصدر: Nature communications. 12(1)

مصطلحات موضوعية: Cornea, NIH 3T3 Cells, Macrophages, Animals, Mice, Knockout, Humans, Mice, Herpesvirus 1, Human, Herpes Simplex, Disease Models, Animal, Tumor Necrosis Factor-alpha, Organ Culture Techniques, Apoptosis, Models, Immunological, Female, Male, Host-Pathogen Interactions, Time-Lapse Imaging, Primary Cell Culture, Intravital Microscopy

الوصف: Rapid death of infected cells is an important antiviral strategy. However, fast decisions that are based on limited evidence can be erroneous and cause unnecessary cell death and subsequent tissue damage. How cells optimize their death decision making strategy to maximize both speed and accuracy is unclear. Here, we show that exposure to TNF, which is secreted by macrophages during viral infection, causes cells to change their decision strategy from "slow and accurate" to "fast and error-prone". Mathematical modeling combined with experiments in cell culture and whole organ culture show that the regulation of the cell death decision strategy is critical to prevent HSV-1 spread. These findings demonstrate that immune regulation of cellular cognitive processes dynamically changes a tissues' tolerance for self-damage, which is required to protect against viral spread.

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الوصول الحر: https://escholarship.org/uc/item/6b9166hkTest

المؤلفون: Jumelle, Clotilde, Sani, Ehsan Shirzaei, Taketani, Yukako, Yung, Ann, Gantin, Fanny, Chauhan, Sunil K, Annabi, Nasim, Dana, Reza

مصطلحات موضوعية: Engineering, Biomedical Engineering, Biotechnology, Physical Injury - Accidents and Adverse Effects, Eye Disease and Disorders of Vision, Eye, Animals, Cornea, Corneal Injuries, Hepatocyte Growth Factor, Hydrogels, Organ Culture Techniques, Swine, Wound Healing, Defect, Hydrogel, Growth factor, Wound healing, Materials Engineering, Biomedical engineering, Materials engineering

الوصف: With 1.5-2.0 million new cases annually worldwide, corneal injury represents a common cause of vision loss, often from irreversible scarring due to surface corneal defects. In this study, we assessed the use of hepatocyte growth factor (HGF) loaded into an in situ photopolymerizable transparent gelatin-based hydrogel for the management of corneal defects. In vitro release kinetics showed that, in regard to the total amount of HGF released over a month, 55 ± 11% was released during the first 24 h, followed by a slow release profile for up to one month. The effect of HGF was assessed using an ex vivo model of pig corneal defect. After three days of organ culture, epithelial defects were found to be completely healed for 89% of the corneas treated with HGF, compared to only 11% of the corneas that had fully re-epithelialized when treated with the hydrogel without HGF. The thickness of the epithelial layer was found to be significantly higher for the HGF-treated group compared to the group treated with hydrogel without HGF (p = 0.0012). Finally, histological and immunostaining assessments demonstrated a better stratification and adhesion of the epithelial layer in the presence of HGF. These results suggest that the HGF-loaded hydrogel system represents a promising solution for the treatment of persistent corneal defects at risk of scarring.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/5c48r32wTest

المؤلفون: Yoon, Paul Sukwoo, Del Piccolo, Nuala, Shirure, Venktesh S, Peng, Yushuan, Kirane, Amanda, Canter, Robert J, Fields, Ryan C, George, Steven C, Gholami, Sepideh

مصطلحات موضوعية: Immunization, Cancer, Colo-Rectal Cancer, Liver Disease, Digestive Diseases, Animals, Cells, Cultured, Colorectal Neoplasms, Disease Models, Animal, Disease Progression, Humans, Immunotherapy, Organ Culture Techniques, Tissue Engineering, Tumor Microenvironment, colorectal cancer, tumor microenvironment, cancer immunology, tissue engineering, organ-on-a-chip, Immunology, Medical Microbiology

الوصف: Colorectal cancer (CRC) is the third most common cancer and second leading cause of cancer-related death in the US. CRC frequently metastasizes to the liver and these patients have a particularly poor prognosis. The infiltration of immune cells into CRC tumors and liver metastases accurately predicts disease progression and patient survival. Despite the evident influence of immune cells in the CRC tumor microenvironment (TME), efforts to identify immunotherapies for CRC patients have been limited. Here, we argue that preclinical model systems that recapitulate key features of the tumor microenvironment-including tumor, stromal, and immune cells; the extracellular matrix; and the vasculature-are crucial for studies of immunity in the CRC TME and the utility of immunotherapies for CRC patients. We briefly review the discoveries, advantages, and disadvantages of current in vitro and in vivo model systems, including 2D cell culture models, 3D culture systems, murine models, and organ-on-a-chip technologies.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/21f5r5qdTest

المؤلفون: Liu, Yizhong, Sakolish, Courtney, Chen, Zunwei, Phan, Duc TT, Bender, R Hugh F, Hughes, Christopher CW, Rusyn, Ivan

مصطلحات موضوعية: Pharmacology and Pharmaceutical Sciences, Biomedical and Clinical Sciences, Cancer, Bioengineering, Biotechnology, 5.1 Pharmaceuticals, 5.9 Resources and infrastructure (treatment development), Development of treatments and therapeutic interventions, Antineoplastic Agents, Cell Culture Techniques, Dose-Response Relationship, Drug, Endothelial Cells, HCT116 Cells, Human Umbilical Vein Endothelial Cells, Humans, Lab-On-A-Chip Devices, Microfluidic Analytical Techniques, Neoplasms, Neovascularization, Pathologic, Organ Culture Techniques, Reproducibility of Results, Endothelial cell, Microphysiological system, Drug safety evaluation, Tissue chip, Toxicology, Pharmacology and pharmaceutical sciences

الوصف: Angiogenesis is a complex process that is required for development and tissue regeneration and it may be affected by many pathological conditions. Chemicals and drugs can impact formation and maintenance of the vascular networks; these effects may be both desirable (e.g., anti-cancer drugs) or unwanted (e.g., side effects of drugs). A number of in vivo and in vitro models exist for studies of angiogenesis and endothelial cell function, including organ-on-a-chip microphysiological systems. An arrayed organ-on-a-chip platform on a 96-well plate footprint that incorporates perfused microvessels, with and without tumors, was recently developed and it was shown that survival of the surrounding tissue was dependent on delivery of nutrients through the vessels. Here we describe a technology transfer of this complex microphysiological model between laboratories and demonstrate that reproducibility and robustness of these tissue chip-enabled experiments depend primarily on the source of the endothelial cells. The model was highly reproducible between laboratories and was used to demonstrate the advantages of the perfusable vascular networks for drug safety evaluation. As a proof-of-concept, we tested Fluorouracil (1-1,000 μM), Vincristine (1-1,000 nM), and Sorafenib (0.1-100 μM), in the perfusable and non-perfusable micro-organs, and in a colon cancer-containing micro-tumor model. Tissue chip experiments were compared to the traditional monolayer cultures of endothelial or tumor cells. These studies showed that human in vitro vascularized micro-organ and micro-tumor models are reproducible organ-on-a-chip platforms for studies of anticancer drugs. The data from the 3D models confirmed advantages of the physiological environment as compared to 2D cell cultures. We demonstrated how these models can be translated into practice by verifying that the endothelial cell source and passage are critical elements for establishing a perfusable model.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/9z99j335Test
https://escholarship.org/content/qt9z99j335/qt9z99j335.pdfTest

المؤلفون: Chen, Hui, Gotimer, Kristin, De Souza, Cristabelle, Tepper, Clifford G, Karnezis, Anthony N, Leiserowitz, Gary S, Chien, Jeremy, Smith, Lloyd H

المصدر: Gynecologic Oncology. 157(3)

مصطلحات موضوعية: Biomedical and Clinical Sciences, Oncology and Carcinogenesis, Ovarian Cancer, Biotechnology, Cancer, Orphan Drug, Rare Diseases, Good Health and Well Being, Aged, Cystadenoma, Serous, Female, Humans, Middle Aged, Neoplasm Grading, Organ Culture Techniques, Organoids, Ovarian Neoplasms, Ovarian cancer, Patient-derived organoid models, Multicellular spheroids, Targeted experimental therapeutics, Paediatrics and Reproductive Medicine, Oncology & Carcinogenesis, Clinical sciences, Oncology and carcinogenesis, Reproductive medicine

الوصف: ObjectiveCancer patient-derived organoids (PDOs) grow as three dimensional (3D) structures in the presence of extracellular matrix and have been found to represent the original tumor's genetic complexity. In addition, PDOs can be grown and subjected to drug sensitivity testing in a shorter time course and with lesser expense than patient-derived xenograft models. Many patients with recurrent ovarian cancer develop malignant effusions that become refractory to chemotherapy. Since these same patients often present for palliative aspiration of ascites or pleural effusions, there is a potential opportunity to obtain tumor specimens in the form of multicellular spheroids (MCS) present in malignant effusion fluids. Our objective was to develop a short duration culture of MCS from ovarian cancer malignant effusions in conditions selected to support organoid growth and use them as a platform for empirical drug sensitivity testing.MethodsIn this study, malignant effusion specimens were collected from patients with high-grade serous ovarian carcinoma (HGSOC). MCS were recovered and subjected to culture conditions designed to support organoid growth. In a subset of specimens, RNA-sequencing was performed at two time points during the short-term culture to determine changes in transcriptome in response to culture conditions. Organoid induction was also characterized in these specimens using Ki67 staining and histologic analysis. Drug sensitivity testing was performed on all specimens.ResultsOur model describes organoids formed within days of primary culture, which can recapitulate the histological features of malignant ascites fluid and can be expanded for at least 6 days. RNA-seq analysis of four patient specimens showed that within 6 days of culture, there was significant up-regulation of genes related to cellular proliferation, epithelial-mesenchymal transition, and KRAS signaling pathways. Drug sensitivity testing identified several agents with therapeutic potential.ConclusionsShort duration organoid culture of MCS from HGSOC malignant effusions can be used as a platform for empiric drug sensitivity testing. These ex vivo models may be helpful in screening new or existing therapeutic agents prior to individualized treatment options.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/3fk3q5nqTest

المؤلفون: Wolf, Kayla J, Shukla, Poojan, Springer, Kelsey, Lee, Stacey, Coombes, Jason D, Choy, Caleb J, Kenny, Samuel J, Xu, Ke, Kumar, Sanjay

المصدر: Proceedings of the National Academy of Sciences of the United States of America. 117(21)

مصطلحات موضوعية: Neurosciences, Biotechnology, 1.1 Normal biological development and functioning, Underpinning research, Actins, Animals, Brain, Cell Adhesion, Cell Line, Tumor, Cell Movement, Extracellular Matrix, Female, Gene Knockout Techniques, Glioblastoma, Humans, Hyaluronan Receptors, Hyaluronic Acid, Mice, Microtubule-Associated Proteins, Microtubules, Myosins, Neoplasm Proteins, Oligopeptides, Organ Culture Techniques, ras GTPase-Activating Proteins, glioblastoma, hyaluronic acid, extracellular matrix, mechanobiology, motility

الوصف: The structure and mechanics of many connective tissues are dictated by a collagen-rich extracellular matrix (ECM), where collagen fibers provide topological cues that direct cell migration. However, comparatively little is known about how cells navigate the hyaluronic acid (HA)-rich, nanoporous ECM of the brain, a problem with fundamental implications for development, inflammation, and tumor invasion. Here, we demonstrate that glioblastoma cells adhere to and invade HA-rich matrix using microtentacles (McTNs), which extend tens of micrometers from the cell body and are distinct from filopodia. We observe these structures in continuous culture models and primary patient-derived tumor cells, as well as in synthetic HA matrix and organotypic brain slices. High-magnification and superresolution imaging reveals McTNs are dynamic, CD44-coated tubular protrusions containing microtubules and actin filaments, which respectively drive McTN extension and retraction. Molecular mechanistic studies reveal that McTNs are stabilized by an interplay between microtubule-driven protrusion, actomyosin-driven retraction, and CD44-mediated adhesion, where adhesive and cytoskeletal components are mechanistically coupled by an IQGAP1-CLIP170 complex. McTNs represent a previously unappreciated mechanism through which cells engage nanoporous HA matrix and may represent an important molecular target in physiology and disease.

وصف الملف: application/pdf

الوصول الحر: https://escholarship.org/uc/item/2fh6t4grTest