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Osteolytic bone lesion is a major cause of lower quality of life and poor prognosis in patients with multiple myeloma (MM), but molecular pathogenesis of the osteolytic process in MM remains elusive. Fms-like tyrosine kinase 3 ligand (FLT3L) was reported to be elevated in bone marrow (BM) and blood of patients with advanced MM who often show osteolysis. Here, we investigated a functional link of FLT3L to osteolytic process in MM. We recruited 86, 306, and 52 patients with MM, acute myeloid leukemia (AML), and acute lymphoblastic leukemia (ALL), respectively. FLT3L levels of patients with hematologic malignancies were measured in BM-derived plasma and found to be significantly higher in MM than in AML or ALL, which rarely show osteolysis. FLT3L levels were further elevated in MM patients with bone lesion compared with patients without bone lesion. In vitro cell-based assays showed that the administration of FLT3L to HEK293T, HeLa, and U2OS cells led to an increase in the DKK1 transcript level through STAT3 phosphorylation at tyrosine 705. WNT reporter assay showed that FLT3L treatment reduced WNT signaling and nuclear translocation of ß-catenin. These results collectively show that the FLT3L-STAT3-DKK1 pathway inhibits WNT signaling-mediated bone formation in MM, which can cause osteolytic bone lesion. Finally, transcriptomic profiles revealed that FLT3L and DKK1 were predominantly elevated in the hyperdiploidy subtype of MM. Taken together, FLT3L can serve as a promising biomarker for predicting osteolytic bone lesion and also a potential therapeutic target to prohibit the progression of the osteolytic process in MM with hyperdiploidy.
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Mieloma Múltiple , Osteólisis , Humanos , Mieloma Múltiple/genética , Mieloma Múltiple/patología , Mieloma Múltiple/metabolismo , Osteólisis/patología , Osteólisis/genética , Osteólisis/etiología , Vía de Señalización Wnt , Masculino , Femenino , Persona de Mediana Edad , Anciano , Línea Celular Tumoral , Factor de Transcripción STAT3/metabolismo , Factor de Transcripción STAT3/genética , Estadificación de Neoplasias , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/metabolismo , AdultoRESUMEN
COVID-19 has infected millions of people worldwide over the past few years. The main technique used for COVID-19 detection is reverse transcription, which is expensive, sensitive, and requires medical expertise. X-ray imaging is an alternative and more accessible technique. This study aimed to improve detection accuracy to create a computer-aided diagnostic tool. Combining other artificial intelligence applications techniques with radiological imaging can help detect different diseases. This study proposes a technique for the automatic detection of COVID-19 and other chest-related diseases using digital chest X-ray images of suspected patients by applying transfer learning (TL) algorithms. For this purpose, two balanced datasets, Dataset-1 and Dataset-2, were created by combining four public databases and collecting images from recently published articles. Dataset-1 consisted of 6000 chest X-ray images with 1500 for each class. Dataset-2 consisted of 7200 images with 1200 for each class. To train and test the model, TL with nine pretrained convolutional neural networks (CNNs) was used with augmentation as a preprocessing method. The network was trained to classify using five classifiers: two-class classifier (normal and COVID-19); three-class classifier (normal, COVID-19, and viral pneumonia), four-class classifier (normal, viral pneumonia, COVID-19, and tuberculosis (Tb)), five-class classifier (normal, bacterial pneumonia, COVID-19, Tb, and pneumothorax), and six-class classifier (normal, bacterial pneumonia, COVID-19, viral pneumonia, Tb, and pneumothorax). For two, three, four, five, and six classes, our model achieved a maximum accuracy of 99.83, 98.11, 97.00, 94.66, and 87.29%, respectively.
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COVID-19 , Aprendizaje Profundo , Neumonía Bacteriana , Neumonía Viral , Neumotórax , Humanos , COVID-19/diagnóstico , SARS-CoV-2 , Inteligencia ArtificialRESUMEN
Walking is an exercise that uses muscles and joints of the human body and is essential for understanding body condition. Analyzing body movements through gait has been studied and applied in human identification, sports science, and medicine. This study investigated a spatiotemporal graph convolutional network model (ST-GCN), using attention techniques applied to pathological-gait classification from the collected skeletal information. The focus of this study was twofold. The first objective was extracting spatiotemporal features from skeletal information presented by joint connections and applying these features to graph convolutional neural networks. The second objective was developing an attention mechanism for spatiotemporal graph convolutional neural networks, to focus on important joints in the current gait. This model establishes a pathological-gait-classification system for diagnosing sarcopenia. Experiments on three datasets, namely NTU RGB+D, pathological gait of GIST, and multimodal-gait symmetry (MMGS), validate that the proposed model outperforms existing models in gait classification.
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Algoritmos , Redes Neurales de la Computación , Marcha , HumanosRESUMEN
Little is known about the ability for epithelial regeneration and wound healing in patients with inflammatory bowel diseases. We evaluated the epithelial proliferation and wound healing ability of patients with Crohn's disease (CD) using patient-derived intestinal organoids. Human intestinal organoids were constructed in a three-dimensional intestinal crypt culture of enteroscopic biopsy samples from controls and CD patients. The organoid-forming efficiency of ileal crypts derived from CD patients was reduced compared with those from control subjects (p < 0.001). Long-term cultured organoids (≥6 passages) derived from controls and CD patients showed an indistinguishable microscopic appearance and culturing behavior. Under TNFα-enriched conditions (30 ng/mL), the organoid reconstitution rate and cell viability of CD patient-derived organoids were significantly lower than those of the control organoids (p < 0.05 for each). The number of EdU+ cells was significantly lower in TNFα-treated organoids derived from CD patients than in TNFα-treated control organoids (p < 0.05). In a wound healing assay, the unhealed area in TNFα-treated CD patient-derived organoids was significantly larger than that of TNFα-treated control organoids (p < 0.001). The wound healing ability of CD patient-derived organoids is reduced in TNFα-enriched conditions, due to reduced cell proliferation. Epithelial regeneration ability may be impaired in patients with CD.
Asunto(s)
Proliferación Celular/genética , Enfermedad de Crohn/terapia , Células Epiteliales/metabolismo , Organoides/crecimiento & desarrollo , Adulto , Enfermedad de Crohn/metabolismo , Enfermedad de Crohn/patología , Células Epiteliales/patología , Femenino , Humanos , Íleon/crecimiento & desarrollo , Íleon/lesiones , Íleon/patología , Mucosa Intestinal/crecimiento & desarrollo , Mucosa Intestinal/patología , Intestinos/diagnóstico por imagen , Intestinos/lesiones , Masculino , Persona de Mediana Edad , Organoides/metabolismo , Regeneración/genética , Transducción de Señal/genética , Células Madre/citología , Células Madre/metabolismo , Factor de Necrosis Tumoral alfa/genética , Cicatrización de Heridas/genéticaRESUMEN
When radiotherapy is applied to the abdomen or pelvis, normal tissue toxicity in the gastrointestinal (GI) tract is considered a major dose-limiting factor. Proton beam therapy has a specific advantage in terms of reduced doses to normal tissues. This study investigated the fundamental differences between proton- and X-ray-induced intestinal injuries in mouse models. C57BL/6J mice were irradiated with 6-MV X-rays or 230-MeV protons and were sacrificed after 84 h. The number of surviving crypts per circumference of the jejunum was identified using Hematoxylin and Eosin staining. Diverse intestinal stem cell (ISC) populations and apoptotic cells were analyzed using immunohistochemistry (IHC) and a terminal deoxynucleotidyl transferase-mediated dUTP nick-end labelling (TUNEL) assay, respectively. The crypt microcolony assay revealed a radiation-dose-dependent decrease in the number of regenerative crypts in the mouse jejunum; proton irradiation was more effective than X-ray irradiation with a relative biological effectiveness of 1.14. The jejunum is the most sensitive to radiations, followed by the ileum and the colon. Both types of radiation therapy decreased the number of radiosensitive, active cycling ISC populations. However, a higher number of radioresistant, reserve ISC populations and Paneth cells were eradicated by proton irradiation than X-ray irradiation, as shown in the IHC analyses. The TUNEL assay revealed that proton irradiation was more effective in enhancing apoptotic cell death than X-ray irradiation. This study conducted a detailed analysis on the effects of proton irradiation versus X-ray irradiation on intestinal crypt regeneration in mouse models. Our findings revealed that proton irradiation has a direct effect on ISC populations, which may result in an increase in the risk of GI toxicity during proton beam therapy.
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Intestinos/lesiones , Protones/efectos adversos , Traumatismos por Radiación/etiología , Rayos X/efectos adversos , Animales , Apoptosis/efectos de la radiación , Modelos Animales de Enfermedad , Relación Dosis-Respuesta en la Radiación , Intestinos/patología , Intestinos/efectos de la radiación , Yeyuno/lesiones , Yeyuno/patología , Yeyuno/efectos de la radiación , Ratones Endogámicos C57BL , Traumatismos por Radiación/patología , Células Madre/patología , Células Madre/efectos de la radiaciónRESUMEN
Nucleotide-binding oligomerization domain-containing protein 2 (NOD2) affords stem cell protection and links microbes to intestinal epithelial regeneration. We investigated whether NOD2 status is associated with crypt survival and intestinal epithelial regeneration independent of microbiota-derived molecules. To assess crypt survival, a clonogenic microcolony assay was performed with 15 Gy of X-ray irradiation. The fractional crypt survival rate (46.0 ± 15.5% vs. 24.7 ± 9.2%, p < 0.01) and fractional EdU-positive crypt survival rate (29.8 ± 14.5% vs. 9.79 ± 4.37%, p = 0.015) were significantly decreased in the NOD2-/- mice compared with the wild-type (WT) mice at 3.5 days after irradiation. To evaluate intestinal epithelial regeneration capability, organoid reconstitution assays were performed. Small bowel crypts of the WT and NOD2-/- mice were isolated and seeded into Matrigel for 3D culture. In the organoid reconstitution assays, the number of organoids formed did not differ between the NOD2-/- and WT mice. Organoid formation ability was also assessed after exposure to 5 Gy irradiation. Organoid formation ability was significantly decreased in the NOD2-/- mice compared with the WT ones after exposure to 5 Gy irradiation (33.2 ± 5.9 vs. 19.7 ± 8.8/well, p < 0.01). NOD2 supports crypt survival after potentially lethal irradiation damage and is associated with intestinal epithelial regeneration.
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Epitelio/patología , Intestinos/patología , Proteína Adaptadora de Señalización NOD2/metabolismo , Traumatismos por Radiación/patología , Regeneración , Animales , Apoptosis/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Células Epiteliales/patología , Células Epiteliales/efectos de la radiación , Ratones Endogámicos C57BL , Proteína Adaptadora de Señalización NOD2/deficiencia , Organoides/patología , Rayos XRESUMEN
The cytidine deaminase (CDA) catalyzes the irreversible hydrolytic deamination of the cytarabine (AraC) into a 1-ß-D-arabinofuranosyluracil (AraU), an inactive metabolite that plays a crucial role in lowering the amount of AraC, a key chemotherapeutic drug, in the treatment of patients with acute myeloid leukemia (AML). In this study, we hypothesized that CDA polymorphisms were associated with the AraC metabolism for AML treatment and/or related clinical phenotypes. We analyzed 16 polymorphisms of CDA among 50 normal karyotype AML (NK-AML) patients, 45 abnormal karyotype AML (AK-AML) patients and 241 normal controls (NC). Several polymorphisms and haplotypes, rs532545, rs2072671, rs471760, rs4655226, rs818194 and CDA-ht3, were found to have a strong correlation with NK-AML compared with NC and these polymorphisms also revealed strong linkage disequilibrium with each other. Among them, rs2072671 (79A>C), which is located in a coding region and the resultant amino acid change K27Q, showed significant associations with NK-AML compared with NC (P=0.009 and odds ratio=2.44 in the dominant model). The AC and CC genotypes of rs2072671 (79A>C) were significantly correlated with shorter overall survival rates (P=0.03, hazard ratio=1.84) and first complete remission duration (P=0.007, hazard ratio=3.24) compared with the AA genotype in the NK-AML patients. Our results indicate that rs2072671 in CDA may be an important prognostic marker in NK-AML patients.
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Citarabina/administración & dosificación , Citidina Desaminasa/genética , Haplotipos , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/genética , Polimorfismo Genético , Adolescente , Adulto , Anciano , Supervivencia sin Enfermedad , Femenino , Humanos , Cariotipo , Leucemia Mieloide Aguda/enzimología , Leucemia Mieloide Aguda/mortalidad , Desequilibrio de Ligamiento , Masculino , Persona de Mediana Edad , Tasa de SupervivenciaRESUMEN
AIMS: The cytosolic 5'-nucleotidase-III (NT5C3) is involved in the metabolism of the nucleoside analog, cytosine arabinose (AraC), and the expression level of NT5C3 is correlated with sensitivity to AraC in acute myeloid leukemia (AML) patients. The current study examined whether the NT5C3 polymorphisms could affect chemotherapy outcomes in 103 Korean AML patients. METHODS: Forty-seven single nucleotide polymorphisms in NT5C3 were genotyped using the Illumina GoldenGate genotyping assay. The genetic effects of the polymorphisms on the outcome of chemotherapy were analyzed using χ and logistic regression models. RESULTS: Although none of the NT5C3 polymorphisms was associated with a complete remission rate, a common single nucleotide polymorphism, rs3750117, showed a significant association with induction rate after the first course of chemotherapy (Pcorr=0.004 and odds ratio=11.28) in AML patients. In addition, NT5C3 expression levels were significantly increased in patients with risk allele homozygote. CONCLUSIONS: The data suggest that genotyping the NT5C3 polymorphism may have the potential to identify patients more likely to respond to AraC-based chemotherapy.
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5'-Nucleotidasa/genética , Antimetabolitos Antineoplásicos/uso terapéutico , Citarabina/uso terapéutico , Glicoproteínas/genética , Quimioterapia de Inducción/métodos , Leucemia Mieloide Aguda/tratamiento farmacológico , Femenino , Estudios de Asociación Genética , Variación Genética , Genotipo , Humanos , Leucemia Mieloide Aguda/genética , Masculino , Polimorfismo de Nucleótido Simple , República de Corea , Resultado del TratamientoRESUMEN
The phantom array effect is one of the temporal light artefacts that can decrease performance and increase fatigue. The phantom array effect visibility shows large individual differences; however, the dominant factors that can explain these individual differences remain unclear. We investigated the relationship between saccadic eye movement speed and phantom array visibility at two different angles and four different directions of saccadic eye movement. The peak speed of saccadic eye movement and the phantom array effect visibility were measured at different modulation frequencies of the light source. Our results show that phantom array visibility increased as eye movement speed increased; the phantom array visibility was higher at a wide viewing angle with fast eye movement speed than at a narrow viewing angle. Moreover, when clustered into subgroups according to individual eye movement speed, the mean speed of the saccadic eye movement of each subgroup is related to the variations in the visibility of the phantom array effect of the subgroup. Therefore, saccadic eye movement speed is related to variations in phantom array effect visibility.
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Movimientos Oculares , Movimientos Sacádicos , Humanos , FatigaRESUMEN
BACKGROUND: Th17 cells and their signature cytokine, interleukin-17A [IL-17], are considered as the main pathogenic factors in inflammatory bowel diseases [IBDs]. However, IL-17 neutralising antibodies, a theoretically curative medication for IBDs, paradoxically aggravated intestinal inflammation. The mechanisms by which IL-17 mediates the protective and pathological effects of IL-17 remain unclear in the intestinal epithelium. METHODS: The intestinal epithelial responses induced by IL-17 were evaluated using the human small intestinal organoid [enteroid] model. RESULTS: Organoid-forming efficiency, cell viability, and proliferation of enteroids were decreased in proportion to IL-17 concentration. The IL-17 induced cytotoxicity was predominantly mediated by pyroptosis with activation of CASP1 and cleavage of GSDMD. Bulk RNA-sequencing revealed the enrichment of secretion signalling in IL-17 treated enteroids, leading to mucin exocytosis. Among its components, PIGR was up-regulated significantly as the concentration of IL-17 increased, resulting in IgA transcytosis. Mucin exocytosis and IgA transcytosis have a protective role against enteric pathogens. Single-cell RNA sequencing identified that CASP1-mediated pyroptosis occurred actively in intestinal stem cells [ISCs] and enterocytes. IL-17 neutralising antibody completely restored IL-17 induced cytotoxicity, but suppressed mucin secretion and IgA transcytosis. Pyroptosis inhibition using CASP1 inhibitors significantly improved IL-17 induced cytotoxicity without diminishing its beneficial effects. CONCLUSIONS: IL-17 induces the pyroptosis of ISCs and enterocytes, as well as mucin secretion of goblet cells and IgA transcytosis of epithelial cells. Paradoxical gastrointestinal effects of IL-17 neutralising antibodies may be associated with inhibition of mucin secretion and IgA transcytosis. The inhibition of pyroptosis using CASP1 inhibitors prevents IL-17 induced cytotoxicity without compromising its beneficial effects.
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Células Madre Adultas , Enfermedades Inflamatorias del Intestino , Adulto , Humanos , Organoides/patología , Interleucina-17/farmacología , Mucosa Intestinal/patología , Mucinas , Enfermedades Inflamatorias del Intestino/patología , Células Madre Adultas/patología , Inmunoglobulina A , Anticuerpos Neutralizantes/farmacologíaRESUMEN
Background: The intestinal microenvironment directly determines the human T-cell receptor (TCR) repertoire. Despite its extreme diversity, TCR repertoire analysis may provide a better understanding of the immune system in patients with inflammatory bowel disease. Methods: To investigate TCR repertoires in the intestinal mucosa, RNA sequencing was performed for inflamed and non-inflamed intestinal mucosa samples obtained from 13 patients with Crohn's disease (CD) and healthy mucosa from nine non-IBD controls. Results: The gene expression frequency of the TCR repertoire showed a clear separation between inflamed mucosa of patients with CD and healthy mucosa of non-IBD controls in the hierarchical clustering heatmap. The richness of TCR repertoires measured by the Chao1 index did not show a significant difference among groups, whereas diversity measured by the D50 diversity index was decreased in the inflamed mucosa of CD patients. Rare/small TCR clonotypes occupied a large proportion of TCR repertoires in healthy mucosa of controls, whereas expanded clonotypes were common in inflamed mucosa of patients with CD. Segment usages of TRAV2, TRAV22, TRAV40, TRJ14, TRAJ51, TRBV1, TRBV21.1, and TRBJ1.5 were significantly decreased in CD patients. KEGG enrichment analysis identified the enrichment of several KEGG pathways, including inflammatory bowel disease (p = 0.0012), Th1 and Th2 cell differentiation (p = 0.0011), and intestinal immune network for IgA production (p = 0.0468). Conclusions: The diversity of the TCR repertoire is reduced in inflamed mucosa of CD patients, which might contribute to intestinal inflammation.
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Enfermedad de Crohn , Enfermedades Inflamatorias del Intestino , Humanos , Mucosa Intestinal , Receptores de Antígenos de Linfocitos TRESUMEN
BACKGROUND & AIMS: Tumor necrosis factor alpha (TNFα) is considered a major tissue damage-promoting effector in Crohn's disease (CD) pathogenesis. Patient-derived intestinal organoid (enteroid) recapitulates the disease-specific characteristics of the intestinal epithelium. This study aimed to evaluate the intestinal epithelial responses to TNFα in enteroids derived from healthy controls and compare them with those of CD patient-derived enteroids. METHODS: Human enteroids derived from patients with CD and controls were treated with TNFα (30 ng/mL), and cell viability and gene expression patterns were evaluated. RESULTS: TNFα induced MLKL-mediated necroptotic cell death, which was more pronounced in CD patient-derived enteroids than in control enteroids. Immunohistochemistry and RNA sequencing revealed that treatment with TNFα caused expansion of the intestinal stem cell (ISC) populations. However, expanded ISC subpopulations differed in control and CD patient-derived enteroids, with LGR5+ active ISCs in control enteroids and reserve ISCs, such as BMI1+ cells, in CD patient-derived enteroids. In single-cell RNA sequencing, LGR5+ ISC-enriched cell cluster showed strong expression of TNFRSF1B (TNFR2) and cyclooxygenase-prostaglandin E2 (PGE2) activation. In TNFα-treated CD patient-derived enteroids, exogenous PGE2 (10 nmol/L) induced the expansion of the LGR5+ ISC population and improved organoid-forming efficiency, viability, and wound healing. CONCLUSIONS: TNFα increases necroptosis of differentiated cells and induces the expansion of LGR5+ ISCs. In CD patient-derived enteroids, TNFα causes LGR5+ stem cell dysfunction (expansion failure), and exogenous PGE2 treatment restored the functions of LGR5+ stem cells. Therefore, PGE2 can be used to promote mucosal healing in patients with CD.
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Enfermedad de Crohn , Factor de Necrosis Tumoral alfa , Enfermedad de Crohn/patología , Humanos , Mucosa Intestinal/patología , Organoides/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Células Madre/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: SAMP1/YitFcsJ (SAMP1) mice spontaneously develop terminal ileitis resembling human Crohn disease. SAMP1 mice have exhibited alteration of epithelial cell lineage distribution and an overall proliferation of the crypt cell population; however, it has not been evaluated whether epithelial differentiation is impaired because of dysfunction of intestinal stem cells (ISCs) or their niche factors. METHODS: Using the intestine of SAMP1 mice aged 10 to 14 weeks, morphometric alterations in the crypt-villus architecture, ISCs, crypt cells, and differentiated cells; organoid formation capacity of intestinal crypts; and niche signaling pathways were analyzed and compared with those of age-matched control AKR/J (AKR) mice. RESULTS: The ileum of SAMP1 mice showed increased depth of intestinal crypts and decreased surface area of the villi compared with those in the ileum of AKR mice. The number of ISCs in the ileal crypts did not differ between SAMP1 and AKR mice; however, the number of Paneth cells decreased and the number of transient amplifying cells increased. The organoid formation rate of the ileal crypts of SAMP1 mice decreased significantly compared with that of AKR mice. The performance of RNA sequencing for intestinal crypts found that the expression of ISC niche factors, such as Wnt3, Dll1, and Dll4, was decreased significantly in the ileal crypts of SAMP1 mice compared with those of AKR mice. Among the ISC niche signals, the Notch signaling-related genes tended to be downregulated. In particular, immunocytochemistry revealed that the expression of Paneth cell-expressing Notch ligand Dll4 was significantly decreased in the intestinal tissue and organoids of SAMP1 mice compared with those of AKR mice. CONCLUSIONS: Depletion of niche factors for ISCs contributes to the alteration of epithelial differentiation in SAMP1 mice.
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Enfermedad de Crohn , Células Epiteliales/citología , Ileítis , Nicho de Células Madre , Animales , Diferenciación Celular , Enfermedad de Crohn/patología , Ileítis/patología , Mucosa Intestinal/citología , Ratones , Ratones EndogámicosRESUMEN
Human renal cell carcinoma (HRCC) is characterized by a high level of resistance to all treatment modalities. Therefore, the investigation of global gene expression in HRCC might help understand its biologic behavior and develop treatment strategies. Using cDNA microarray analysis, we initially compared gene expression profiles between HRCCs and adjacent normal tissues, and found that 87 were up-regulated and 127 genes were down-regulated. Next, a subset of genes, twofold differentially expressed, were validated by Northern blotting. Unexpectedly, caveolin-1, a gene reported to be a tumor suppressor gene, was found to be up-regulated in HRCC tissues. Expression level of caveolin-1 in SN12CPM6 (high metastatic clone) was higher than in SN12C (low metastatic clone), and SN12CPM6 was more resistant to doxorubicin (DXR) than SN12C. Caveolin-1 gene was slightly induced in surviving SN12C cells after DXR treatment. Furthermore, SN12CPM6-siCav1 cells, which were transfected with siRNA of cavelon-1 gene, were more sensitive to DXR, compared to SN12CPM6, but reduction of caveolin-1 gene expression did not affect tumor formation in subcapsule of kidney and lung metastasis. On the other hand, induction of caveolin-1 gene affected the production of lung metastasis under anti-cancer drug treatment: the incidence of pulmonary metastasis was significantly lower in SCID mice injected with SN12CPM6-siCav1 cells, and the number of pulmonary nodules decreased significantly (p = 0.0004). The above results together suggest that caveolin-1 may confer a growth advantage to cancer cells during DXR chemotherapy and surviving HRCC cells eventually might develop lung metastasis.
Asunto(s)
Apoptosis/efectos de los fármacos , Carcinoma de Células Renales/patología , Caveolina 1/metabolismo , Doxorrubicina/farmacología , Neoplasias Renales/patología , Neoplasias Pulmonares/prevención & control , Interferencia de ARN/fisiología , Animales , Antibióticos Antineoplásicos/farmacología , Carcinoma de Células Renales/metabolismo , Caveolina 1/genética , Línea Celular Tumoral , Regulación hacia Abajo/efectos de los fármacos , Humanos , Neoplasias Renales/metabolismo , Neoplasias Pulmonares/secundario , Ratones , Ratones Desnudos , Ratones SCID , Metástasis de la Neoplasia/prevención & control , ARN Interferente Pequeño/genética , Transfección , Trasplante Heterólogo , Regulación hacia Arriba/efectos de los fármacosRESUMEN
BACKGROUND/AIMS: The modulation of CD1d-restricted natural killer T (NKT) cells by glycolipids has been considered as a potential therapy against immunologic diseases, including inflammatory bowel disease. A recently identified a glycolipid analog, 7DW8-5, which is derived from α-galactosylceramide (α-GalCer), is as much as 100-fold more active at stimulating both human and mice NKT cells when compared to α-GalCer. We explored the effects of 7DW8-5 in mouse models of acute and chronic colitis. METHODS: We investigated the effects of 7DW8-5 on intestinal inflammation by assessing the effects of 7dW8-5 on a murine dextran sulfate sodium (DSS)-induced acute colitis model and a chronic colitis-associated tumor model. RESULTS: The acute DSS-induced colitis model showed a dose-dependent response to 7DW8-5, as mice administered 7DW8-5 showed a significant improvement in DSS-induced colitis based on their disease activity index, histologic analysis, and serum C-reactive protein levels, when compared to mice administered vehicle alone. However, DSS-induced colitis in CD1d-KO mice showed no response to 7DW8-5. A fluorescence-activating cell sorting analysis revealed an increase in NKT cells in colonic tissues of 7DW8-5-treated mice. RNA-seq and real-time quantitative polymerase chain reaction showed a significant increase in the expression of interleukin (IL)-4, IL-13, and interferon-gamma in 7DW8-5-treated mice. In addition, 7DW8-5 treatment reduced colitis-associated tumor development in an azoxymethane/DSS mouse model. CONCLUSIONS: 7DW8-5 activates NKT cells through CD1d and provides a protective effect against intestinal inflammation in mice. Therefore, 7DW8-5 may be a promising therapeutic agent for treatment of inflammatory bowel disease.
RESUMEN
Cancer cells exhibit properties of cells in a less differentiated state than the adjacent normal cells in the tissue. We explored whether cancer cells can be converted to a differentiated normal-like state by restoring the gene regulatory network (GRN) of normal cells. Here, we report that colorectal cancer cells exhibit a range of developmental states from embryonic and intestinal stem-like cells to differentiated normal-like cells. To identify the transcription factors (TF) that commit stem-like colorectal cancer cells into a differentiated normal-like state, we reconstructed GRNs of normal colon mucosa and identified core TFs (CDX2, ELF3, HNF4G, PPARG, and VDR) that govern the cellular state. We further found that SET Domain Bifurcated 1 (SETDB1), a histone H3 lysine 9-specific methyltransferase, hinders the function of the identified TFs. SETDB1 depletion effectively converts stem-like colorectal cancer cells into postmitotic cells and restores normal morphology in patient-derived colorectal cancer organoids. RNA-sequencing analyses revealed that SETDB1 depletion recapitulates global gene expression profiles of normal differentiated cells by restoring the transcriptional activity of core TFs on their target genes. IMPLICATIONS: Our study provides insights into the molecular regulatory mechanism underlying the developmental hierarchy of colorectal cancer and suggests that induction of a postmitotic state may be a therapeutic alternative to destruction of cancer cells.
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Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , N-Metiltransferasa de Histona-Lisina/genética , Células CACO-2 , Diferenciación Celular/fisiología , Línea Celular Tumoral , Neoplasias Colorrectales/metabolismo , Células Madre Embrionarias/patología , Regulación Neoplásica de la Expresión Génica , Redes Reguladoras de Genes , Células HCT116 , N-Metiltransferasa de Histona-Lisina/metabolismo , Humanos , Células Madre Neoplásicas/patología , Transfección , Células Tumorales CultivadasRESUMEN
The administration of a combination of probiotics and prebiotics is expected to be a promising strategy for improving irritable bowel syndrome (IBS) symptoms. This study aimed to investigate the efficacy of a synbiotic containing Lactobacillus paracasei and Opuntia humifusa extract for symptomatic improvement of IBS in a murine model and to evaluate the mechanism underlying the beneficial effects of this synbiotic. A total of 20 male Wistar rats aged 8 weeks with IBS induced by restraint stress were assigned into four groups and administered L. paracasei as a probiotic and O. humifusa extract as a prebiotic for 4 weeks. The primary outcome was stool consistency at week 4. To evaluate the mechanism underlying the beneficial effects of the synbiotic, fecal microbial analysis was conducted, and the serum corticosterone levels, tumor necrosis factor-α (TNF-α) levels in the colon tissue, and expression of tight junction proteins were investigated. All three treatment groups showed significantly lower scores for stool consistency than the control group at week 4 (all p < 0.001). When compared with the control group, the synbiotic groups showed a significantly greater abundance of L. paracasei in fecal microbial analysis, lower serum corticosterone levels, lower TNF-α levels in the colon tissue, and higher expression of tight junction proteins. This novel synbiotic containing L. paracasei and O. humifusa extract can improve the stool consistency in a murine model of IBS. It may be a promising treatment option for IBS, and human studies are warranted.
Asunto(s)
Síndrome del Colon Irritable/terapia , Lacticaseibacillus paracasei/fisiología , Opuntia/química , Extractos Vegetales/administración & dosificación , Simbióticos/administración & dosificación , Animales , Colon/química , Corticosterona/sangre , Modelos Animales de Enfermedad , Heces/microbiología , Masculino , Prebióticos/administración & dosificación , Probióticos/administración & dosificación , Ratas , Ratas Wistar , Proteínas de Uniones Estrechas/análisisRESUMEN
Background/Aims: Dysbiosis is an important factor in the pathogenesis of irritable bowel syndrome (IBS). Several studies have reported promising results using probiotics for the treatment of IBS. This study evaluated the efficacy of novel probiotics isolated from Kimchi, a Korean fermented food, and the feces of healthy Vietnamese people in a murine model of IBS. Methods: Lactobacillus paracasei DK121 was isolated from Kimchi, and L. salivarius V4 and L. plantarum V7 were isolated from the feces of healthy Vietnamese people residing in Korea. Forty rats were allocated to receive one of the study strains, a mixture of the strains, or the vehicle. After 5 days of administration, the rats were restrained in a cage to induce IBS. The effects of the probiotics on IBS were analyzed by evaluating the stool weights and stool consistency scores. Results: The primary outcome was analyzed upon the completion of a three-week experiment. The rats in the V7 group showed lower stool weights than those in the control group at week 2 (median: 1.10 [V7] vs. 2.35 [control], p=0.04, Mann-Whitney U-test) and week 3 (median: 1.10 [V7] vs. 2.80 [control], p=0.017). The rats in the DK121 (median: 2.00, p=0.007), V7 (median: 2.00, p=0.004), and mixture (median: 1.50, p=0.001) groups showed better stool consistency scores at week 2 than the control group (median: 3.00). Conclusions: The novel probiotics have beneficial effects on defecation in a murine model of IBS. Human studies confirming the efficacy are warranted.
Asunto(s)
Síndrome del Colon Irritable/terapia , Probióticos/administración & dosificación , Animales , Peso Corporal , Modelos Animales de Enfermedad , Heces/microbiología , Humanos , Síndrome del Colon Irritable/patología , Lactobacillus/aislamiento & purificación , Lactobacillus/fisiología , Masculino , Ratas , Ratas WistarRESUMEN
We present a framework for monocular 3D kinematic pose tracking and viewpoint estimation of periodic and quasi-periodic human motions from an uncalibrated camera. The approach we introduce here is based on learning both the visual observation manifold and the kinematic manifold of the motion using a joint representation. We show that the visual manifold of the observed shape of a human performing a periodic motion, observed from different viewpoints, is topologically equivalent to a torus manifold. The approach we introduce here is based on the supervised learning of both the visual and kinematic manifolds. Instead of learning an embedding of the manifold, we learn the geometric deformation between an ideal manifold (conceptual equivalent topological structure) and a twisted version of the manifold (the data). Experimental results show accurate estimation of the 3D body posture and the viewpoint from a single uncalibrated camera.
Asunto(s)
Algoritmos , Interpretación de Imagen Asistida por Computador/métodos , Imagenología Tridimensional/métodos , Movimiento , Reconocimiento de Normas Patrones Automatizadas/métodos , Postura , Imagen de Cuerpo Entero/métodos , Inteligencia Artificial , Humanos , Aumento de la Imagen/métodos , Modelos Biológicos , Modelos Estadísticos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Técnica de SustracciónRESUMEN
AIMS: NOD2 and CD1d play a key role in innate immunity by recognizing conserved molecular patterns of pathogens. While NOD2-/- and CD1d-/- mice display structural and functional alterations in Paneth cells, animal studies have reported no impact of NOD2 or CD1d deficiency on experimental colitis. NOD2 mutations increase the susceptibility to inflammatory bowel diseases and the CD1d bound to α-galactosylceramide [α-GalCer] alleviates intestinal inflammation. We evaluated the effect of CD1d modulation on experimental colitis in NOD2-/- mice. METHODS: The effect of CD1d augmentation and depletion in NOD2-/- mice was assessed in a dextran sodium sulphate [DSS]-induced colitis model via administration of α-GalCer and construction of NOD2-/-CD1d-/- mice. The structural and functional changes in Paneth cells were evaluated using transmission electron microscopy and pilocarpine administration. Colitogenic taxa were analysed in the faeces of NOD2-/-CD1d-/- mice using 16S rRNA gene sequencing. RESULTS: In NOD2-/- mice, α-GalCer alleviated and CD1d depletion [NOD2-/-CD1d-/- mice] aggravated colitis activity and histology compared with co-housed littermates NOD2-/-, CD1d-/- and wild-type mice after administration of 3% DSS. In NOD2-/-CD1d-/- mice, the ultrastructure and degranulation ability of secretary granules in Paneth cells were altered and the intestinal microbial composition differed from that of their littermates. Faecal microbiota transplantation [FMT] with NOD2-/-CD1d-/- mice faeces into wild-type mice aggravated DSS-induced colitis, while FMT with wild-type mice faeces into NOD2-/-CD1d-/- mice alleviated DSS-induced colitis. Acetatifactor muris was identified only in NOD2-/-CD1d-/- mice faeces and the oral gavage of A. muris in wild-type mice aggravated DSS-induced colitis. CONCLUSION: CD1d modulates colonic inflammation in NOD2-/- mice by altering the intestinal microbial composition comprising A. muris.