Kidney organoids derived from human pluripotent stem cells (hPSCs) have extensive potential for disease modelling and regenerative medicine. However, the limited vascularization and immaturity of ...kidney organoids have been still remained to overcome. Extracellular matrix (ECM) can provide mechanical support and a biochemical microenvironment for cell growth and differentiation. Here in vitro methods using a kidney decellularized extracellular matrix (dECM) hydrogel to culture hPSC‐derived kidney organoids, which have extensive vascular network and their own endothelial cells, are reported. Single‐cell transcriptomics reveal that the vascularized kidney organoids cultured using the kidney dECM have more mature patterns of glomerular development and higher similarity to human kidney than those cultured without the kidney dECM. Differentiation of α‐galactosidase A (GLA)‐knock‐out hPSCs generated using CRISPR/Cas9 into kidney organoids by the culture method using kidney dECM efficiently recapitulate Fabry nephropathy with vasculopathy. Transplantation of kidney organoids with kidney dECM into kidney of mouse accelerates the recruitment of endothelial cells from the host mouse kidney and maintains vascular integrity with the more organized slit diaphragm‐like structures than those without kidney dECM. The kidney dECM methodology for inducing extensive vascularization and maturation of kidney organoids can be applied to studies for kidney development, disease modeling, and regenerative medicine.
The limited vascularization and immaturity of kidney organoids are still to be overcome. Here in vitro methods using a kidney dECM hydrogel to culture hPSC‐derived kidney organoids, which have extensive vascular network with their own endothelial cells and matured nephron‐like structures, are reported. This methodology can be applied to studies for kidney development, disease modeling, and regenerative medicine.
Accurate prediction of treatment response for cancer patients is essential for overcoming intrinsic therapy resistance that results from genetic heterogeneity, varying tumor growth kinetics, and the ...complex tumor microenvironment. To achieve this goal, there is an urgent need for effective preclinical in vitro models that recapitulate the molecular–pathologic features and intricate ecology of native tumors for precision medicine. In this study, a vascularized organoid model (VOM) composed of patient‐derived gastric cancer organoids (PDOs), perfusable endothelium, and stomach decellularized extracellular matrix is presented that enables the prediction of clinical response to VEGFR2‐targeted therapy in gastric cancer patients. The results indicate that VOMs are dependent on the PDO molecular subtype. Moreover, VOMs accurately reproduce the clinically observed responses of patients treated with VEGFR2 inhibitor. Therefore, VOMs represent a valuable platform for providing clinical predictions for personalized testing and potential discovery of therapeutic drugs in various cancers that lack standardized regimens.
The Vascularized Organoid Model (VOM) accurately reproduces the drug response observed in patients in a clinic by providing a highly biomimetic cancer system to PDOs. The system is composed of stomach‐specific and collagen‐enriched materials, interstitial‐like tissue, and a perfusable blood vessel that enables drug delivery through vessels to the organoids instead of direct exposure.
Tremendous advances have been made toward accurate recapitulation of the human intestinal system in vitro to understand its developmental process, and disease progression. However, current in vitro ...models are often confined to 2D or 2.5D microarchitectures, which is difficult to mimic the systemic level of complexity of the native tissue. To overcome this problem, physiologically relevant intestinal models are developed with a 3D hollow tubular structure using 3D bioprinting strategy. A tissue‐specific biomaterial, colon‐derived decellularized extracellular matrix (Colon dECM) is developed and it provides significant maturation‐guiding potential to human intestinal cells. To fabricate a perfusable tubular model, a simultaneous printing process of multiple materials through concentrically assembled nozzles is developed and a light‐activated Colon dECM bioink is employed by supplementing with ruthenium/sodium persulfate as a photoinitiator. The bioprinted intestinal tissue models show spontaneous 3D morphogenesis of the human intestinal epithelium without any external stimuli. In consequence, the printed cells form multicellular aggregates and cysts and then differentiate into several types of enterocytes, building junctional networks. This system can serve as a platform to evaluate the effects of potential drug‐induced toxicity on the human intestinal tissue and create a coculture model with commensal microbes and immune cells for future therapeutics.
A robust bioprinting strategy for the fabrication of tubular intestine models is developed using light‐activated tissue‐specific biomaterials. The colon‐derived decellularized extracellular matrix bioink is supplemented with the photoinitiator ruthenium/sodium persulfate (Ru/SPS) to enhance the shape fidelity during the printing process. The bioprinted intestine model shows accelerated maturation of intestinal cells and 3D epithelial morphogenesis.
Adipose tissue is highly vascularized and requires the angiogenic properties for its mass growth. Visfatin has been recently characterized as a novel adipokine, which is preferentially produced by ...adipose tissue. In this study, we report that visfatin potently stimulates in vivo neovascularization in chick chorioallantoic membrane and mouse Matrigel plug. We also demonstrate that visfatin activates migration, invasion, and tube formation in human umbilical vein endothelial cells (HUVECs). Moreover, visfatin evokes activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) in endothelial cells, which is closely linked to angiogenesis. Inhibition of ERK activation markedly decreases visfatin-induced tube formation of HUVECs and visfatin-stimulated endothelial cell sprouting from rat aortic rings. Taken together, these results demonstrate that visfatin promotes angiogenesis via activation of mitogen-activated protein kinase ERK-dependent pathway and suggest that visfatin may play important roles in various pathophysiological angiogenesis including adipose tissue angiogenesis.
Non‐neural extracellular matrix (ECM) has limited application in humanized physiological neural modeling due to insufficient brain‐specificity and safety concerns. Although brain‐derived ECM contains ...enriched neural components, certain essential components are partially lost during the decellularization process, necessitating augmentation. Here, it is demonstrated that the laminin‐augmented porcine brain‐decellularized ECM (P‐BdECM) is xenogeneic factor‐depleted as well as favorable for the regulation of human neurons, astrocytes, and microglia. P‐BdECM composition is comparable to human BdECM regarding brain‐specificity through the matrisome and gene ontology‐biological process analysis. As augmenting strategy, laminin 111 supplement promotes neural function by synergic effect with laminin 521 in P‐BdECM. Annexin A1(ANXA1) and Peroxiredoxin(PRDX) in P‐BdECM stabilized microglial and astrocytic behavior under normal while promoting active neuroinflammation in response to neuropathological factors. Further, supplementation of the brain‐specific molecule to non‐neural matrix also ameliorated glial cell inflammation as in P‐BdECM. In conclusion, P‐BdECM‐augmentation strategy can be used to recapitulate humanized pathophysiological cerebral environments for neurological study.
Porcine‐derived brain decellularized extracellular matrix (P‐BdECM) augmented with LN111 is developed to construct physiological human brain model. The similarity of P‐BdECM is specifically validated to human BdECM and brain‐specific proteins in ameliorating neural differentiation and neuroinflammation. The augmented P‐BdECM is favorable to recapitulate the neural microenvironment for the study of cellular behavior and the role of ECM in neural processes.
A wide variety of experimental models including 2D cell cultures, model organisms, and 3D
in vitro
models have been developed to understand pathophysiological phenomena and assess the safety and ...efficacy of potential therapeutics. In this sense, 3D
in vitro
models are an intermediate between 2D cell cultures and animal models, as they adequately reproduce 3D microenvironments and human physiology while also being controllable and reproducible. Particularly, recent advances in 3D
in vitro
biomimicry models, which can produce complex cell structures, shapes, and arrangements, can more similarly reflect
in vivo
conditions than 2D cell culture. Based on this, 3D bioprinting technology, which enables to place the desired materials in the desired locations, has been introduced to fabricate tissue models with high structural similarity to the native tissues. Therefore, this review discusses the recent developments in this field and the key features of various types of 3D-bioprinted tissues, particularly those associated with blood vessels or highly vascularized organs, such as the heart, liver, and kidney. Moreover, this review also summarizes the current state of the three categories: (1) chemical substance treatment, (2) 3D bioprinting of lesions, and (3) recapitulation of tumor microenvironments (TME) of 3D bioprinting-based disease models according to their disease modeling approach. Finally, we propose the future directions of 3D bioprinting approaches for the creation of more advanced
in vitro
biomimetic 3D tissues, as well as the translation of 3D bioprinted tissue models to clinical applications.
Gaucher disease (GD) is caused by a deficiency of β-glucocerebrosidase, encoded by GBA. Haplotype analyses previously demonstrated founder effects for particular GBA mutations in Ashkenazi Jewish and ...French-Canadian populations. This study aimed to investigate the clinical characteristics and mutation spectrum of GBA in Korean GD patients and to identify founder effect of GBA p.G85E in non-neuronopathic GD patients.
The study cohort included 62 GD patients from 58 unrelated families. Among them, 18 patients from 17 families harbored the p.G85E mutation. Haplotype analysis was performed for 9 probands and their parents for whom DNA samples were available. In 58 unrelated probands, the GBA mutation p.L483P was the most common (30/116 alleles, 26%), followed by p.G85E (16%), p.F252I (13%), and p.R296Q (9%). The median age at diagnosis of the 18 patients harboring the p.G85E mutation was 3.8 (range 1.2-57) years. No patients developed neurological symptoms during follow-up periods of 2.2-20.3 (median 13.9) years. The size of the shared haplotype containing GBA p.G85E was 732 kbp, leading to an estimated age of 3075 years.
The GBA p.G85E mutation, which appears to be neuroprotective despite producing distinctive visceromegaly and skeletal symptoms, exhibited a potential founder effect in Korean GD patients.
Recently, respiratory systems are increasingly threatened by high levels of environmental pollution. Organ-on-a-chip technology has the advantage of enabling more accurate preclinical experiments by ...reproducing in vivo organ physiology. To investigate disease mechanisms and treatment options, respiratory-physiology-on-a-chip systems have been studied for the last decade. Here, we delineate the strategic approaches to develop respiratory-physiology-on-a-chip that can recapitulate respiratory system in vitro. The state-of-the-art biofabrication methods and biomaterials are considered as key contributions to constructing the chips. We also explore the vascularization strategies to investigate complicated pathophysiological phenomena including inflammation and immune responses, which are the critical aggravating factors causing the complications in the respiratory diseases. In addition, challenges and future research directions are delineated to improve the mimicry of respiratory systems in terms of both structural and biological behaviors.
Despite encouraging progress in the development of
cancer models,
cancer models that simultaneously recapitulate the complexity of the tumor microenvironment and its diverse cellular components and ...genetic properties remain lacking. Here, an advanced vascularized lung cancer (LC) model is proposed, which includes patient-derived LC organoids (LCOs), lung fibroblasts, and perfusable vessels using 3D bioprinting technology. To better recapitulate the biochemical composition of native lung tissues, a porcine lung-derived decellularized extracellular matrix (LudECM) hydrogel was produced to offer physical and biochemical cues to cells in the LC microenvironment. In particular, idiopathic pulmonary fibrosis-derived lung fibroblasts were used to implement fibrotic niches similar to actual human fibrosis. It was shown that they increased cell proliferation and the expression of drug resistance-related genes in LCOs with fibrosis. In addition, changes in resistance to sensitizing targeted anti-cancer drugs in LCOs with fibrosis were significantly greater in LudECM than in that Matrigel. Therefore, assessment of drug responsiveness in vascularized LC models that recapitulate lung fibrosis can help determine the appropriate therapy for LC patients accompanied by fibrosis. Furthermore, it is expected that this approach could be utilized for the development of targeted therapies or the identification of biomarkers for LC patients accompanied by fibrosis.
The switch/sucrose nonfermenting (SWI/SNF) complex is an adenosine triphosphate-dependent chromatin-remodeling complex associated with the regulation of DNA accessibility. Germline mutations in the ...components of the SWI/SNF complex are related to human developmental disorders, including the Coffin-Siris syndrome (CSS), Nicolaides-Baraitser syndrome (NCBRS), and nonsyndromic intellectual disability. These disorders are collectively referred to as SWI/SNF complex-related intellectual disability disorders (SSRIDDs).
Whole-exome sequencing was performed in 564 Korean patients with neurodevelopmental disorders. Twelve patients with SSRIDDs (2.1%) were identified and their medical records were retrospectively analyzed.
ARID1B, found in eight patients, was the most frequently altered gene. Four patients harbored pathogenic variants in SMARCA4, SMARCB1, ARID2, and SMARCA2. Ten patients were diagnosed with CSS, and one patient without a typical phenotype was diagnosed with ARID1B-related nonsyndromic intellectual disability. Another patient harboring the SMARCA2 pathogenic variant was diagnosed with NCBRS. All pathogenic variants in ARID1B were truncating, whereas variants in SMARCA2, SMARCB1, and SMARCA4 were nontruncating (missense). Frequently observed phenotypes were thick eyebrows (10/12), hypertrichosis (8/12), coarse face (8/12), thick lips (8/12), and long eyelashes (8/12). Developmental delay was observed in all patients, and profound speech delay was also characteristic. Agenesis or hypoplasia of the corpus callosum was observed in half of the patients (6/12).
SSRIDDs have a broad disease spectrum, including NCBRS, CSS, and ARID1B-related nonsyndromic intellectual disability. Thus, SSRIDDs should be considered as a small but important cause of human developmental disorders.
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