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Los Organoides (O) son un tipo de cultivo celular 3D, que reproducen las características morfológicas y funcionales de diversos órganos o tejidos en un entorno in vivo. Se logran a través de la proliferación y diferenciación de Células Madres (CM) en distintas líneas celulares con capacidad de autoorganizarse. Son capaces de reproducir forma, función, expresión génica o repuesta a estímulos de la misma forma que el órgano original. Esto le ha permitido servir de base para múltiples investigaciones en el ámbito médico y odontológico. En los últimos años, se ha podido recrear con éxito, prácticamente, todos los órganos de nuestro cuerpo, como pulmones, hígado, tracto reproductivo, cerebro y muchos otros (Bartfeld, 2021). De la misma forma, son varias las líneas de investigación odontológicas desarrolladas. En específico, la creación de O de órganos orales como dientes y glándulas salivales, son las más reportadas (Oshima et al., 2017). Sin embargo, no son del común conocimiento del odontólogo general. Esta revisión sistemática exploratoria, tiene como objetivo presentar una visión general de la evidencia acumulada, determinado las áreas odontológicas de investigación, así como sus resultados. La investigación odontológica, en base al uso de O, es de alta calidad y de vanguardia, mostrando resultados prometedores, que auguran un gran futuro, tanto para la odontología como para los pacientes.
Organoids (O) are a type of 3D cell culture, which reproduce the morphological and functional characteristics of various organs or tissues in an in vivo environment. They are achieved through the proliferation and differentiation of Stem Cells (SC) into different cell lines with the ability to self-organize. They are capable of reproducing form, function, gene expression, or responses to stimuli in the same way as the original organ. This has allowed it to serve as the basis for multiple investigations in the medical and dental field. In recent years, it has been possible to successfully recreate practically all human organs, such as the lungs, liver, reproductive tract, brain and many others (Bartfeld, 2021). In the same way, there are several lines of dental research developed, specifically, the creation of O from oral organs such as teeth and salivary glands, are the most reported (Oshima et al., 2017). However, they are not common knowledge of the general dentist. This exploratory systematic review aims to present an overview of the accumulated evidence, determining the dental research areas, as well as their results. Dental research, based on the use of O, is of high quality and cutting-edge, showing promising results and a favorable future, both for dentistry and for patients.
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The hepatocellular carcinoma (HCC) features a remarkable epidemiological burden, ranking as the third most lethal cancer worldwide. As the HCC-related molecular and cellular complexity unfolds as the disease progresses, the use of a myriad of in vitro models available is mandatory in translational preclinical research setups. In this review paper, we will compile cutting-edge information on the in vitro bioassays for HCC research, (A) emphasizing their morphological and molecular parallels with human HCC; (B) delineating the advantages and limitations of their application; and (C) offering perspectives on their prospective applications. While bidimensional (2D) (co) culture setups provide a rapid low-cost strategy for metabolism and drug screening investigations, tridimensional (3D) (co) culture bioassays - including patient-derived protocols as organoids and precision cut slices - surpass some of the 2D strategies limitations, mimicking the complex microarchitecture and cellular and non-cellular microenvironment observed in human HCC. 3D models have become invaluable tools to unveil HCC pathophysiology and targeted therapy. In both setups, the recapitulation of HCC in different etiologies/backgrounds (i.e., viral, fibrosis, and fatty liver) may be considered as a fundamental guide for obtaining translational findings. Therefore, a "multimodel" approach - encompassing the advantages of different in vitro bioassays - is encouraged to circumvent "model-biased" outcomes in preclinical HCC research.
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Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/genética , Animais , Carcinogênese/patologia , Carcinogênese/genética , Organoides/patologia , Modelos BiológicosRESUMO
Considering the importance of alternative methodologies to animal experimentation, we propose an organoid-based biological model for in vitro blood vessel generation, achieved through co-culturing endothelial and vascular smooth muscle cells (VSMCs). Initially, the organoids underwent comprehensive characterization, revealing VSMCs (α-SMA + cells) at the periphery and endothelial cells (CD31+ cells) at the core. Additionally, ephrin B2 and ephrin B4, genes implicated in arterial and venous formation respectively, were used to validate the obtained organoid. Moreover, the data indicates exclusive HIF-1α expression in VSMCs, identified through various methodologies. Subsequently, we tested the hypothesis that the generated blood vessels have the capacity to modulate the osteogenic phenotype, demonstrating the ability of HIF-1α to promote osteogenic signals, primarily by influencing Runx2 expression. Overall, this study underscores that the methodology employed to create blood vessel organoids establishes an experimental framework capable of producing a 3D culture model of both venous and arterial endothelial tissues. This model effectively guides morphogenesis from mesenchymal stem cells through paracrine signaling, ultimately leading to an osteogenic acquisition phenotype, with the dynamic involvement of HIF-1α.
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Subunidade alfa do Fator 1 Induzível por Hipóxia , Músculo Liso Vascular , Miócitos de Músculo Liso , Organoides , Osteogênese , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Osteogênese/genética , Organoides/metabolismo , Organoides/citologia , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/citologia , Miócitos de Músculo Liso/metabolismo , Miócitos de Músculo Liso/citologia , Células Cultivadas , Vasos Sanguíneos/metabolismo , Vasos Sanguíneos/citologia , Vasos Sanguíneos/crescimento & desenvolvimento , Técnicas de Cocultura/métodos , Diferenciação Celular , Células Endoteliais/metabolismo , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologiaRESUMO
This review delves into the groundbreaking impact of induced pluripotent stem cells (iPSCs) and three-dimensional organoid models in propelling forward neuropathology research. With a focus on neurodegenerative diseases, neuromotor disorders, and related conditions, iPSCs provide a platform for personalized disease modeling, holding significant potential for regenerative therapy and drug discovery. The adaptability of iPSCs, along with associated methodologies, enables the generation of various types of neural cell differentiations and their integration into three-dimensional organoid models, effectively replicating complex tissue structures in vitro. Key advancements in organoid and iPSC generation protocols, alongside the careful selection of donor cell types, are emphasized as critical steps in harnessing these technologies to mitigate tumorigenic risks and other hurdles. Encouragingly, iPSCs show promising outcomes in regenerative therapies, as evidenced by their successful application in animal models.
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Células-Tronco Pluripotentes Induzidas , Organoides , Organoides/patologia , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Animais , Neuropatologia/métodos , Medicina Regenerativa/métodos , Doenças Neurodegenerativas/terapia , Doenças Neurodegenerativas/patologia , Diferenciação CelularRESUMO
Three-dimensional (3D) bioprinting, a promising advancement in tissue engineering technology, involves the robotic, layer-by-layer additive biofabrication of functional 3D tissue and organ constructs. This process utilizes biomaterials, typically hydrogels and living cells, following digital models. Traditional tissue engineering uses a classic triad of living cells, scaffolds, and physicochemical signals in bioreactors. A scaffold is a temporary, often biodegradable, support structure. Tissue engineering primarily falls into two categories: (i) scaffold based and (ii) scaffold free. The latter, scaffold-free 3D bioprinting, is gaining increasing popularity. Organ building blocks (OBB), capable of self-assembly and self-organization, such as tissue spheroids, organoids, and assembloids, have begun to be utilized in scaffold-free bioprinting. This article discusses the expanding range of OBB, presents the rapidly evolving collection of bioprinting and bioassembly methods using these OBB, and finally, outlines the advantages, challenges, and future perspectives of using OBB in organ printing.
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Purpose: This study aimed to compare the degree of maturation and development of fetal pig segmental intestinal tissue with that of spheroids created by in-vitro reaggregation of dissociated fetal intestinal cells after transplantation into immunodeficient mice. Methods: Fetal pig small intestines were transplanted as segmental grafts into the omentum and subrenal capsules of immunodeficient mice or enzymatically treated to generate single cells. Spheroids made by in-vitro reaggregation of these cells were transplanted into the subrenal capsules of immunodeficient mice. The segmental grafts and spheroids were harvested four and eight weeks after transplantation, and the structural maturity and in-vivo development of these specimens were histologically evaluated. Results: The spheroids were engrafted and supplied blood vessels from the host mice, but an intestinal layered structure was not clearly observed, and there was almost no change in size. On the other hand, the segmental grafts formed deep crypts in the mucus membrane, the inner circular layer, and outer longitudinal muscles. The crypts of the transplanted grafts harvested at eight weeks were much deeper, and the smooth muscle layer and the enteric nervous system were more mature than those of grafts harvested at the fourth week, although the intestinal peristaltic wave was not observed. Conclusions: Spheroids created from fetal small intestinal cells could not form layered structures or mature sufficiently. Conversely, segmental tissues structurally matured and developed after in-vivo transplantation and are therefore potential grafts for transplantation.
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Animais , Camundongos , Suínos , Transplante Heterólogo/veterinária , Transplante de Tecido Fetal/veterinária , Maturidade dos Órgãos FetaisRESUMO
Advances in the development of pharmacological treatment in oral cancer require tumor models capable of simulating the complex biology of the tumor microenvironment. The spread of three-dimensional models has changed the scenery of in vitro cell culture techniques, contributing to translational oncology. Still, the full extent of their application in preclinical drug trials is yet to be understood. Therefore, the present scoping review protocol was established to screen the literature on using three-dimensional cell culture models in drug-testing assays in the context of oral cancer. This scoping review will be conducted based on the guidelines established by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Extension for Scoping Review guidelines (PRISMA-ScR). We will search the PubMed/Medline, Web of Science, Scopus, and Embase databases, as well as the gray literature, including peer-reviewed research articles involving 3D models applied to drug-assessment assays in oral cancer published from 1 March 2013 until 1 March 2023. Data will be charted, and findings will be described according to the predetermined questions of interest. We will present these findings in a narrative manner.
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Introducción: La pandemia causada por SARS-CoV-2 ha impactado al mundo gravemente en diversos ámbitos y con ello ha surgido la necesidad de contar con herramientas con mayor relevancia fisiológica para investigar patologías complejas como el COVID-19. Los organoides son un modelo experimental con características únicas como la capacidad de autoformar una estructura tridimensional utilizando células en cultivo. Sobre esta base, surge la siguiente pregunta ¿son los organoides un modelo experimental factible para reflejar la fisiopatología del COVID-19 y evaluar la eficacia de fármacos que limiten su progresión? Metodología: Para abordar esta interrogante, esta revisión plantea el analizar la validez de los organoides como modelo experimental y verificar su utilidad en la evaluación de fármacos para el COVID-19. Para cumplir estos objetivos se realizó una revisión sistemática cualitativa de la literatura, a través de una búsqueda en PubMed con el término 'COVID-19 and stem cells and organoids' y también en un número especial de la revista Cell. Resultados: Se organizaron los resultados relevantes por sistema fisiológico y en la evaluación de fármacos. Los organoides más empleados para estudios de COVID-19 correspondieron a tejido respiratorio, nervioso y digestivo. Algunos resultados encontrados en la revisión fueron similares a aquellos obtenidos a partir de tejidos de pacientes COVID-19 o autopsias, encontrándose hallazgos relevantes como la posible disrupción de la barrera epitelial del sistema nervioso por infección del plexo coroideo. También se logró observar efectividad de fármacos que posteriormente pasaron a ser aprobados y utilizados exitosamente en pacientes. Conclusión: Los organoides se pueden componer a partir de diferentes tipos celulares y bajo diferentes protocolos experimentales, siendo relevante la lectura crítica de los artículos científicos para decidir si sus resultados son extrapolables a la fisiopatología de la enfermedad.
Introduction: The pandemic caused by SARS-CoV-2 has impacted the world severely in several aspects and has created the need for research tools to study the COVID-19 disease. Organoids are experimental models with unique characteristics, like the ability to self-assemble in a tridimensional structure. Based on this, the following question arises: are organoids an experimental model suitable to reflect the physiopathology of COVID-19 and to allow the evaluation of the efficacy of drugs that limit its progression? Methods: To approach this question, this review aimed to analyze the validity of organoids as an experimental model and verify their utility in COVID-19 drug evaluation. To resolve these objectives, a qualitative systematic review was done through a PubMed search with the terms 'COVID-19 and stem cells and organoids' and on a special issue of the Cell Journal. Results: The results were organized by physiologic system and therapeutic drug evaluation. The most utilized tissues for the COVID-19 study were respiratory, nervous, and digestive. Some results found in the review were like those obtained from COVID-19 patient tissue or autopsies, finding some relevant discoveries like the possibility of the choroid plexus disruption in the nervous system caused by the infection. Efficacy was also observed in approved drugs and used later in patients successfully. Conclusion: Organoids might be composed starting with different cell types and under a variety of experimental protocols, being relevant the critical reading of the scientific literature to decide whether their results can be extrapolated to the pathophysiology of the disease
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The immune and endocrine dysfunctions of white adipose tissue are a hallmark of metabolic disorders such as obesity and type 2 diabetes. In humans, white adipose tissue comprises distinct depots broadly distributed under the skin (hypodermis) and as internal depots (visceral). Depot-specific ASCs could account for visceral and subcutaneous adipose tissue properties, by regulating adipogenesis and immunomodulation. More importantly, visceral and subcutaneous depots account for distinct contributions to obesity and its metabolic comorbidities. Recently, distinct ASCs subpopulations were also described in subcutaneous adipose tissue. Interestingly, the superficial layer closer to the dermis shows hyperplastic and angiogenic capacities, whereas the deep layer is considered as having inflammatory properties similar to visceral. The aim of this focus review is to bring the light of recent discoveries into white adipose tissue heterogeneity together with the biology of distinct ASCs subpopulations and to explore adipose tissue 3D models revealing their advantages, disadvantages, and contributions to elucidate the role of ASCs in obesity development. Recent advances in adipose tissue organoids opened an avenue of possibilities to recreate the main cellular and molecular events of obesity leading to a deep understanding of this inflammatory disease besides contributing to drug discovery. Furthermore, 3D organ-on-a-chip will add reproducibility to these adipose tissue models contributing to their translation to the pharmaceutical industry.
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Diabetes Mellitus Tipo 2 , Humanos , Reprodutibilidade dos Testes , Diabetes Mellitus Tipo 2/metabolismo , Tecido Adiposo/metabolismo , Gordura Subcutânea , Obesidade/metabolismoRESUMO
Chagas disease (CD) is a life-threatening illness caused by the parasite Trypanosoma cruzi (T. cruzi). With around seven million people infected worldwide and over 50,000 deaths per year, CD is a major public health issue in Latin America. The main route of transmission to humans is through a triatomine bug (vector-borne), but congenital and oral transmission have also been reported. The acute phase of CD presents mild symptoms but may develop into a long-lasting chronic illness, characterized by severely impaired cardiac, digestive, and neurological functions. The intestinal tissue appears to have a key role during oral transmission and chronic infection of CD. In this immune-privileged reservoir, dormant/quiescent parasites have been suggested to contribute to disease persistence, infection relapse, and treatment failure. However, the interaction between the intestinal epithelium and T. cruzi has not been examined in depth, in part, due to the lack of in vitro models that approximate to the biological and structural complexity of this tissue. Therefore, to understand the role played by the intestinal tissue during transmission and chronic infection, physiological models resembling the organ complexity are needed. Here we addressed this issue by establishing and characterizing adult stem cell-derived colonoid infection models that are clinically relevant for CD. 3D and 2D systems of murine intestinal organoids infected with T. cruzi Dm28c (a highly virulent strain associated with oral outbreaks) were analyzed at different time points by confocal microscopy. T. cruzi was able to invade and replicate in intestinal epithelial primary cells grown as intact organoids (3D) and monolayers (2D). The permissiveness to pathogen infection differed markedly between organoids and cell lines (primate and intestinal human cell lines). So far, this represents the first evidence of the potential that these cellular systems offer for the study of host-pathogen interactions and the discovery of effective anti-chagasic drugs.
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Doença de Chagas , Trypanosoma cruzi , Humanos , Animais , Camundongos , Infecção Persistente , Doença de Chagas/parasitologia , Mucosa Intestinal , Colo , OrganoidesRESUMO
Autism spectrum disorders (ASD) are a group of complex neurodevelopmental disorders that affect communication and social interactions and present with restricted interests and repetitive behavior patterns. The susceptibility to ASD is strongly influenced by genetic/heritable factors; however, there is still a large gap in understanding the cellular and molecular mechanisms underlying the neurobiology of ASD. Significant progress has been made in identifying ASD risk genes and the possible convergent pathways regulated by these gene networks during development. The breakthrough of cellular reprogramming technology has allowed the generation of induced pluripotent stem cells (iPSCs) from individuals with syndromic and idiopathic ASD, providing patient-specific cell models for mechanistic studies. In the past decade, protocols for developing brain organoids from these cells have been established, leading to significant advances in the in vitro reproducibility of the early steps of human brain development. Here, we reviewed the most relevant literature regarding the application of brain organoids to the study of ASD, providing the current state of the art, and discussing the impact of such models on the field, limitations, and opportunities for future development.
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Transtorno do Espectro Autista , Células-Tronco Pluripotentes Induzidas , Humanos , Transtorno do Espectro Autista/genética , Reprodutibilidade dos Testes , Encéfalo , OrganoidesRESUMO
Down syndrome (DS, or trisomy 21, T21), is the most common genetic cause of intellectual disability. Alterations in the complex process of cerebral cortex development contribute to the neurological deficits in DS, although the underlying molecular and cellular mechanisms are not completely understood. Human cerebral organoids (COs) derived from three-dimensional (3D) cultures of induced pluripotent stem cells (iPSCs) provide a new avenue for gaining a better understanding of DS neuropathology. In this study, we aimed to generate iPSCs from individuals with DS (T21-iPSCs) and euploid controls using urine-derived cells, which can be easily and noninvasively obtained from most individuals, and examine their ability to differentiate into neurons and astrocytes grown in monolayer cultures, as well as into 3D COs. We employed nonintegrating episomal vectors to generate urine-derived iPSC lines, and a simple-to-use system to produce COs with forebrain identity. We observed that both T21 and control urine-derived iPSC lines successfully differentiate into neurons and astrocytes in monolayer, as well as into COs that recapitulate early features of human cortical development, including organization of neural progenitor zones, programmed differentiation of excitatory and inhibitory neurons, and upper-and deep-layer cortical neurons as well as astrocytes. Our findings demonstrate for the first time the suitability of using urine-derived iPSC lines to produce COs for modeling DS.
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Cérebro , Síndrome de Down , Células-Tronco Pluripotentes Induzidas , Neurogênese , Organoides , Células-Tronco Pluripotentes Induzidas/citologia , Organoides/citologia , Organoides/crescimento & desenvolvimento , Cérebro/citologia , Cérebro/crescimento & desenvolvimento , Síndrome de Down/genética , Síndrome de Down/patologia , Síndrome de Down/urina , Técnicas de Cultura de Células em Três Dimensões , Humanos , Neurônios/citologia , Astrócitos/citologia , Linhagem da CélulaRESUMO
Alzheimer's disease (AD) is the primary cause of dementia, to date. The urgent need to understand the biological and biochemical processes related to this condition, as well as the demand for reliable in vitro models for drug screening, has led to the development of novel techniques, among which stem cell methods are of utmost relevance for AD research, particularly the development of human brain organoids. Brain organoids are three-dimensional cellular aggregates derived from induced pluripotent stem cells (iPSCs) that recreate different neural cell interactions and tissue characteristics in culture. Here, we describe the protocol for the generation of brain organoids derived from AD patients and for the analysis of AD-derived pathology. AD organoids can recapitulate beta-amyloid and tau pathological features, making them a promising model for studying the molecular mechanisms underlying disease and for in vitro drug testing.
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Doença de Alzheimer , Células-Tronco Pluripotentes Induzidas , Humanos , Organoides , Doença de Alzheimer/patologia , Encéfalo/patologia , Peptídeos beta-Amiloides/metabolismoRESUMO
Há muitos anos a cultura celular bidimensional (2D) é utilizada como modelo de estudo de doenças, possuindo grande importância na medicina regenerativa, apesar de ainda conter limitações significativas. A fim de contornar essas limitações, a cultura celular tridimensional (3D) propõe uma organização mais complexa e sustentável que pode ser produzida a partir de células-tronco adultas (ASCs), células-tronco embrionárias (ESCs) ou células-tronco pluripotentes induzidas (iPSCs). A cultura 3D possibilitou o cultivo de células em um ambiente mais próximo do fisiológico, levando à formação de distintos tecidos órgãos-específicos. Em outras palavras, a cultura de células 3D possibilita a criação de estruturas orgânicas muito semelhantes aos órgãos de um ser humano, tanto estruturalmente, quanto funcionalmente. Desse modo, tem-se o que é chamado de organoides. O uso dos organoides tem crescido exponencialmente em ambientes in vitro, permitindo a análise e observação dos diversos fenômenos fisiológicos existentes. Como exemplo, pode-se citar os organoides cerebrais ("mini-brains") reproduzidos in vitro buscando delinear as peculiaridades e complexidades do cérebro humano, com o objetivo de compreender algumas disfunções neurológicas que acometem esse sistema, como as duas principais doenças neurodegenerativas: Doenças de Alzheimer e Parkinson. Portanto, os organoides cerebrais podem permitir notável avanço da medicina regenerativa aplicada a doenças neurodegenerativas, já que esses "mini-brains" podem ser produzidos a partir de células do próprio paciente. Isso permitirá intervenções personalizadas, como testagens farmacológicas, a fim de definir qual seria o melhor tratamento medicamentoso. Consequentemente, essa tecnologia pode permitir terapias mais eficientes e individualizadas - o que é fundamental para a Medicina Personalizada (AU).
For many years, two-dimensional (2D) cell culture has been used as a model to study diseases, having great importance in regenerative medicine, despite still having significant limitations. In order to circumvent these limitations, three-dimensional (3D) cell culture proposes a more complex and sustainable organization that can be produced from adult stem cells (ASCs), embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs). The 3D culture enabled the cultivation of cells in an environment closer to the physiological one, leading to the formation of different organ-specific tissues. In other words, 3D cell culture makes it possible to create organic structures very similar to the organs of a human being, both structurally and functionally. In this way, we have what are called organoids. The use of organoids has grown exponentially in in vitro environments, allowing the analysis and observation of the various existing physiological phenomena. As an example, we can mention the brain organoids ("mini-brains") reproduced in vitro, seeking to delineate the peculiarities and complexities of the human brain, in order to understand some neurological dysfunctions that affect this system, such as the two main neurodegenerative diseases: Alzheimer's and Parkinson's Diseases. Therefore, brain organoids may allow a remarkable advance in regenerative medicine applied to neurodegenerative diseases, as these "mini-brains" can be produced from the patient's own cells. This will allow for personalized interventions, such as drug testing, in order to define what would be the best drug treatment. Consequently, this technology can enable more efficient and individualized therapies - which is fundamental for Personalized Medicine (AU).
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Humanos , Doença de Parkinson , Organoides , Medicina ConciergeRESUMO
Tumor organoids are defined as self-organized three-dimensional assemblies of heterogeneous cell types derived from patient samples that mimic the key histopathological, genetic, and phenotypic characteristics of the original tumor. This technology is proposed as an ideal candidate for the evaluation of possible therapies against cancer, presenting advantages over other models which are currently used. However, there are no reports in the literature that relate the techniques and material development of tumor organoids or that emphasize in the physicochemical and biological properties of materials that intent to biomimicry the tumor extracellular matrix. There is also little information regarding the tools to identify the correspondence of native tumors and tumoral organoids (tumoroids). Moreover, this paper relates the advantages of organoids compared to other models for drug evaluation. A growing interest in tumoral organoids has arisen from 2009 to the present, aimed at standardizing the process of obtaining organoids, which more accurately resemble patient-derived tumor tissue. Likewise, it was found that the characteristics to consider for the development of organoids, and therapeutic responses of them, are cell morphology, physiology, the interaction between cells, the composition of the cellular matrix, and the genetic, phenotypic, and epigenetic characteristics. Currently, organoids have been used for the evaluation of drugs for brain, lung, and colon tumors, among others. In the future, tumor organoids will become closer to being considered a better model for studying cancer in clinical practice, as they can accurately mimic the characteristics of tumors, in turn ensuring that the therapeutic response aligns with the clinical response of patients.
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BACKGROUND: Schizophrenia is a complex and severe neuropsychiatric disorder, with a wide range of debilitating symptoms. Several aspects of its multifactorial complexity are still unknown, and some are accepted to be an early developmental deficiency with a more specifically neurodevelopmental origin. Understanding the timepoints of disturbances during neural cell differentiation processes could lead to an insight into the development of the disorder. In this context, human brain organoids and neural cells differentiated from patient-derived induced pluripotent stem cells are of great interest as a model to study the developmental origins of the disease. RESULTS: Here we evaluated the differential expression of proteins of schizophrenia patient-derived neural progenitors (NPCs), early neurons, and brain organoids in comparison to healthy individuals. Using bottom-up shotgun proteomics with a label-free approach for quantitative analysis, we found multiple dysregulated proteins since NPCs, modified, and disrupted the 21DIV neuronal differentiation, and cerebral organoids. Our experimental methods have shown impairments in pathways never before found in patient-derived induced pluripotent stem cells studies, such as spliceosomes and amino acid metabolism; but also, those such as axonal guidance and synaptogenesis, in line with postmortem tissue studies of schizophrenia patients. CONCLUSION: In conclusion, here we provide comprehensive, large-scale, protein-level data of different neural cell models that may uncover early events in brain development, underlying several of the mechanisms within the origins of schizophrenia.
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The therapeutic use of classical psychedelic substances such as d-lysergic acid diethylamide (LSD) surged in recent years. Studies in rodents suggest that these effects are produced by increased neural plasticity, including stimulation of the mTOR pathway, a key regulator of metabolism, plasticity, and aging. Could psychedelic-induced neural plasticity be harnessed to enhance cognition? Here we show that LSD treatment enhanced performance in a novel object recognition task in rats, and in a visuo-spatial memory task in humans. A proteomic analysis of human brain organoids showed that LSD affected metabolic pathways associated with neural plasticity, including mTOR. To gain insight into the relation of neural plasticity, aging and LSD-induced cognitive gains, we emulated the experiments in rats and humans with a neural network model of a cortico-hippocampal circuit. Using the baseline strength of plasticity as a proxy for age and assuming an increase in plasticity strength related to LSD dose, the simulations provided a good fit for the experimental data. Altogether, the results suggest that LSD has nootropic effects.
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Alucinógenos , Nootrópicos , Animais , Alucinógenos/toxicidade , Humanos , Dietilamida do Ácido Lisérgico/farmacologia , Proteômica , Ratos , Serina-Treonina Quinases TORRESUMO
This thematic issue of Experimental Biology and Medicine is dedicated to the incredibly important contributions made by women leaders in the biomedical sciences throughout recent history. Scientists from many disciplines have contributed papers, both original research and state of the art reviews, to demonstrate the type of work being performed every day by women leaders committed to advancing scientific knowledge in their respective fields of specialization. In this introduction, we provide readers with a brief highlight of the information to be found in the invited papers.
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Pesquisa Biomédica , Medicina , Mulheres , Feminino , Humanos , LiderançaRESUMO
Os sarcomas de partes moles (SPM) são tumores malignos derivados de células mesenquimais, com grande capacidade de formar metástases e geralmente apresentam elevada resistência à terapia. Formam um grupo extremamente heterogêneo com mais de 100 subtipos tumorais, como os sarcomas pleomórficos indiferenciados (SPI) e os sarcomas fusocelulares (SF). Em conjunto, esses dois subgrupos não compartilham semelhanças morfológicas com os outros subtipos. Adicionalmente, carecem de informações moleculares que auxiliem tanto no diagnóstico quanto no prognóstico da doença. O objetivo deste trabalho foi estabelecer e caracterizar modelos pré-clínicos de SPI e SF baseados em Patient-derived xenografts (PDX) e organoids (PDO) para definir uma plataforma de estudos destes tumores e avaliar o potencial de alvos moleculares específicos. Utilizando amostras de 11 pacientes, foi possível estabelecer oito PDX de SPI ou SF. O estabelecimento de organóides de sarcoma se mostrou desafiador para amostras tumorais oriundas diretamente de pacientes ou de PDX e após testarmos diferentes protocolos, conseguimos obter organóides de culturas primárias tumorais. Os organoides e os PDXs gerados foram caracterizados por IHQ e foi possível observar a preservação das características moleculares. Para a caracterização genética, realizamos o sequenciamento de exoma de um conjunto de oito amostras (paciente/PDX/cultura celular) para identificação das principais alterações somáticas. Por fim, padronizamos o ensaio de citotoxicidade à doxorrubicina em três culturas primárias de SPI e SF.
Soft tissue sarcomas (STS) are malignant tumors derived from mesenchymal cells, with a great capacity to form metastases and generally show high resistance to therapy. They form an extremely heterogeneous group with more than 100 tumor subtypes, such as undifferentiated pleomorphic sarcoma (UPS) and fusocellular sarcomas (SS). Taken together, these two subgroups do not share morphological similarities with the other subtypes. Additionally, they lack molecular information that helps both in the diagnosis and prognosis of the disease. The objective of this work was to establish and characterize preclinical models of UPS and SS based on Patient-derived xenografts (PDX) and organoids (PDO) to define a platform for studies of these tumors and to evaluate the potential of specific molecular targets. Using samples from eleven patients, it was possible to establish eight PDX of UPS and SS. The establishment of sarcoma organoids proved to be challenging for tumor samples derived directly from patients or from PDX and after testing different protocols, we were able to obtain organoids from primary tumor cultures. The generated organoids and PDXs were also characterized by IHC and immunofluorescence with sarcoma marker proteins and it was also possible to observe the preservation of molecular characteristics. For genetic characterization, we performed exome sequencing of a set of eight samples (patient/PDX/cell culture) to identify the main somatic alterations. Finally, we standardized the doxorubicin cytotoxicity assay in three primary cultures of SPI and SF.