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1.
Medicine (Baltimore) ; 98(45): e17799, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31702632

RESUMO

Alpha-B crystallin (CRYAB), as a small heat shock protein, has been found to be highly expressed in various human cancers and significantly associated with the unfavorable prognosis of these tumor. Nevertheless, the clinical significance of CRYAB in gastric cancer (GC) angiogenesis remains to be elucidated. In this study, we evaluated the expression of CRYAB and CD34 in GC tissues and corresponding normal gastric specimens to explore whether high level CRYAB is related with the angiogenesis and the poor prognosis in GC.In this study, the expression of CRYAB and CD34 were detected in GC tissues and corresponding normal gastric tissues by immunohistochemical (IHC) technique. Furthermore, the relationship of CRYAB with CD34-evaluated microvessel density (MVD) and poor prognosis was also investigated.CRYAB expression level was significantly higher in GC tissue than in normal gastric mucosa tissue, and clearly mean higher MVD was observed in tumor tissues compared with non-cancerous tissues. Besides, higher MVD value was observed in positive CRYAB expression group than in negative CRYAB expression group. Statistical analysis showed that CRYAB and MVD are associated with clinicopathological features including lymph node metastasis (LNM), tumor differentiation, invasion depth, and TNM stages. Kaplan-Meier method and multivariate survival analysis indicated that high expression of CRYAB, MVD, invasion depth, TNM stages, and tumor differentiation, as well as LNM significantly correlate with poor prognosis of GC patients.High expression of CRYAB may contribute to angiogenesis, invasion and metastasis of GC. These results indicated that CRYAB was expected to be a promising molecular marker for poor prognosis and potential therapeutic target in patients with GC.


Assuntos
Antígenos CD34/metabolismo , Neoplasias Gástricas/patologia , Regulação para Cima , Cadeia B de alfa-Cristalina/metabolismo , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática , Masculino , Invasividade Neoplásica , Estadiamento de Neoplasias , Prognóstico , Neoplasias Gástricas/metabolismo , Análise de Sobrevida
2.
Cancer Sci ; 110(11): 3424-3433, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31495050

RESUMO

Tongue squamous cell carcinoma (TSCC) has a poor prognosis due to its early metastasis through blood and lymphatic vessels. We undertook a systematic review to investigate the prognostic significance of blood microvessel density (MVD) and lymphatic vessel density (LVD) in TSCC patients. We carried out a systematic search in Ovid Medline, Scopus, and Cochrane libraries. All studies that evaluated the prognostic significance of MVD/LVD markers in TSCC were systematically retrieved. Our results showed that MVD/LVD markers, CD31, CD34, CD105, factor VIII, lymphatic vessel endothelial hyaluronan receptor-1, and D2-40 were evaluated in TSCC patients until 28 June 2018. Six out of 13 studies reported markers that were associated with poor prognosis in TSCC. Two out of three studies suggested that a high number of D2-40+ vessels predicated low overall survival (OS); the third study reported that the ratio of D2-40+ over factor VIII+ vessels is associated with low OS. Most of the other markers had controversial results for prognostication. We found higher expression of MVD/LVD markers were commonly, but not always, associated with shorter survival in TSCC patients. It is therefore not currently possible to recommend implementation of these markers as reliable prognosticators in clinical practice. More studies (especially for D2-40) with larger patient cohorts are needed.


Assuntos
Biomarcadores Tumorais/metabolismo , Carcinoma de Células Escamosas/metabolismo , Vasos Linfáticos/metabolismo , Microvasos/metabolismo , Neoplasias da Língua/metabolismo , Anticorpos Monoclonais Murinos/metabolismo , Antígenos CD34/metabolismo , Carcinoma de Células Escamosas/mortalidade , Carcinoma de Células Escamosas/patologia , Endoglina/metabolismo , Fator VIII/metabolismo , Humanos , Receptores de Hialuronatos/metabolismo , Linfangiogênese , Neovascularização Patológica , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Prognóstico , Neoplasias da Língua/mortalidade , Neoplasias da Língua/patologia , Proteínas de Transporte Vesicular/metabolismo
3.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 41(4): 512-516, 2019 Aug 30.
Artigo em Chinês | MEDLINE | ID: mdl-31484614

RESUMO

To study the clinicopathologic characteristics,immunohistochemical features,differential diagnosis,and prognosis of solitary fibrous tumours(SFT)/hemangiopericytomas(HPC)in the maters(meninx). Methods A series of 7 cases previously diagnosed as SFT/HPC at the Department of Pathology,Peking Union Medical College Hospital,during the period from 2008 to 2018 were analyzed for clinical data,histopathology,and immunohistochemical findings.The patients were followed up and the relevant literatures were reviewed. Results These seven patients included two males and 5 females aged 22 to 77 years(mean,49 years).Headache was the most common symptom.The magnetic resonance imaging of SFT/HPC showed irregularly contoured masses and dural tail sign was observed at the periphery of the lesion in 4 cases.The major axis of the tumor ranged from 1.8 cm to 10 cm(mean,4 cm).The tumors were located in the mater in 6 cases and in the spinal meninx in 1 case.The tumors were surgically removed in all cases.Under light microscope,the tumors were formed by long round,oval or spindle cells,with rich branching vascular pattern and varying quantity of collagenous fibers bands in both sparse areas and dense areas.According the WHO classification,2 cases were in WHO grade Ⅰ,2 cases in WHO grade Ⅱ,and 3 cases in WHO grade Ⅲ.Immunohistochemistry of the paraffin-embedded tissues in all cases showed positive immunoreativity for CD34 and vimentin in all seven cases,along with positive signal transducer and activator of transcription 6 in 4 cases,negative epithelial membrane antigen and S-100 in 7 cases,and negative progestational hormone and somatostatin receptor 2 in 6 cases.The Ki-67 index ranged from 1% to 15%.Five patients with follow-up data(including 1 current case)were alive,while 2 patients were lost to follow-up. Conclusions The SFT/HPC are rare in the maters(meninx)and is clinically difficult to be differentiated from other meningioma.The combination of CD34 and signal transducer and activator of transcription 6 helps to diagnose this disease.


Assuntos
Hemangiopericitoma/diagnóstico , Meninges/patologia , Tumores Fibrosos Solitários/diagnóstico , Adulto , Idoso , Antígenos CD34/metabolismo , Diagnóstico Diferencial , Feminino , Hemangiopericitoma/patologia , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Prognóstico , Fator de Transcrição STAT6/metabolismo , Tumores Fibrosos Solitários/patologia , Adulto Jovem
4.
Pan Afr Med J ; 33: 134, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31558933

RESUMO

Primary angiosarcoma of the breast is an extremely rare tumour with a difficult diagnosis and poor prognosis. We report a case of primary breast angiosarcoma diagnosed in the pathology department of the University Hospital of Oujda. An analysis of the epidemiological, diagnostic and therapeutic aspects of this type of tumour is made in this manuscript. Mastectomy is the standard treatment; the place of radiotherapy and chemotherapy is not well established. We report a case of a 18- year-old woman having an infectious symptomatology of the right breast for which she received an anti-infectious therapy inducing regression of inflammatory symptoms presented with a quick growing mass. Initial core needle biopsy showed a malignant vascular proliferation. The patient underwent a mastectomy. The tumor histology showed papillary formations and vascular structures lined by atypical cells with hyperchromatic nucleus and eosinophilic cytoplasm. The tumor cells expressed CD34 and CD31 but were negative for cytokeratin. The diagnosis of angiosarcoma grade I was made. The patient is now receiving chemotherapy. She is still alive.


Assuntos
Neoplasias da Mama/diagnóstico , Hemangiossarcoma/diagnóstico , Mastectomia/métodos , Adolescente , Antígenos CD34/metabolismo , Antineoplásicos/administração & dosagem , Biópsia com Agulha de Grande Calibre , Neoplasias da Mama/patologia , Neoplasias da Mama/cirurgia , Feminino , Hemangiossarcoma/patologia , Hemangiossarcoma/cirurgia , Humanos , Gradação de Tumores , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo
5.
Plast Reconstr Surg ; 144(2): 372-384, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31348346

RESUMO

BACKGROUND: We investigated expression of prorenin receptor, angiotensin-converting enzyme, angiotensin II receptor 1, and angiotensin II receptor 2 by the embryonic stem cell-like population on the endothelium of the microvessels and perivascular cells within keloid-associated lymphoid tissues. METHODS: Immunohistochemical staining for prorenin receptor, angiotensin-converting enzyme, angiotensin II receptor 1, and angiotensin II receptor 2 was performed on 11 formalin-fixed, paraffin-embedded sections of keloid tissue samples. Immunofluorescence staining was performed on three keloid tissue samples by co-staining with OCT4, CD34, ERG, and tryptase. Real-time quantitative polymerase chain reaction was performed on five keloid tissue samples and four keloid-derived primary cell lines. Western blotting was performed on the four keloid-derived primary cell lines for mRNA and protein expression of these proteins, respectively. RESULTS: Immunohistochemical and immunofluorescence staining showed expression of prorenin receptor, angiotensin-converting enzyme, angiotensin II receptor 1, and angiotensin II receptor 2 in all 11 keloid tissue samples. Prorenin receptor and angiotensin II receptor 1 were expressed on the endothelium and the pericyte layer of the microvessels and perivascular cells, angiotensin II receptor 2 was localized to the endothelium of the microvessels and the tryptase-positive perivascular cells, and angiotensin-converting enzyme was localized to the endothelium of the microvessel, within the keloid-associated lymphoid tissues. Real-time quantitative polymerase chain reaction showed transcripts of prorenin receptor, angiotensin-converting enzyme, and angiotensin II receptor 1 in the keloid tissue samples and keloid-derived primary cell lines, whereas angiotensin II receptor 2 was detected in keloid tissue samples only. Western blotting confirmed the presence of prorenin receptor, angiotensin-converting enzyme, and angiotensin II receptor 1 in the keloid-derived primary cell lines. CONCLUSION: Prorenin receptor, angiotensin-converting enzyme, angiotensin II receptor 1, and angiotensin II receptor 2 were expressed by the embryonic stem cell-like population within the keloid-associated lymphoid tissues, suggesting that this primitive population may be a potential therapeutic target by modulation of the renin-angiotensin system.


Assuntos
Células-Tronco Embrionárias/metabolismo , Queloide/metabolismo , Sistema Renina-Angiotensina/fisiologia , Adolescente , Adulto , Antígenos CD34/metabolismo , Linhagem Celular , Criança , Pré-Escolar , Feminino , Imunofluorescência , Humanos , Queloide/patologia , Masculino , Pessoa de Meia-Idade , Fator 3 de Transcrição de Octâmero/metabolismo , Peptidil Dipeptidase A/metabolismo , RNA Mensageiro/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Receptor Tipo 2 de Angiotensina/metabolismo , Receptores de Superfície Celular/metabolismo , Regulador Transcricional ERG/metabolismo , Adulto Jovem
6.
Nature ; 571(7765): 355-360, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31270458

RESUMO

Defining the transcriptomic identity of malignant cells is challenging in the absence of surface markers that distinguish cancer clones from one another, or from admixed non-neoplastic cells. To address this challenge, here we developed Genotyping of Transcriptomes (GoT), a method to integrate genotyping with high-throughput droplet-based single-cell RNA sequencing. We apply GoT to profile 38,290 CD34+ cells from patients with CALR-mutated myeloproliferative neoplasms to study how somatic mutations corrupt the complex process of human haematopoiesis. High-resolution mapping of malignant versus normal haematopoietic progenitors revealed an increasing fitness advantage with myeloid differentiation of cells with mutated CALR. We identified the unfolded protein response as a predominant outcome of CALR mutations, with a considerable dependency on cell identity, as well as upregulation of the NF-κB pathway specifically in uncommitted stem cells. We further extended the GoT toolkit to genotype multiple targets and loci that are distant from transcript ends. Together, these findings reveal that the transcriptional output of somatic mutations in myeloproliferative neoplasms is dependent on the native cell identity.


Assuntos
Genótipo , Mutação , Transtornos Mieloproliferativos/genética , Transtornos Mieloproliferativos/patologia , Neoplasias/genética , Neoplasias/patologia , Transcriptoma/genética , Animais , Antígenos CD34/metabolismo , Calreticulina/genética , Linhagem Celular , Proliferação de Células , Células Clonais/classificação , Células Clonais/metabolismo , Células Clonais/patologia , Endorribonucleases/metabolismo , Hematopoese/genética , Células-Tronco Hematopoéticas/classificação , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Hematopoéticas/patologia , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Camundongos , Modelos Moleculares , Transtornos Mieloproliferativos/classificação , NF-kappa B/metabolismo , Neoplasias/classificação , Células-Tronco Neoplásicas/citologia , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/patologia , Mielofibrose Primária/genética , Mielofibrose Primária/patologia , Proteínas Serina-Treonina Quinases/metabolismo , Análise de Sequência de RNA/métodos , Análise de Célula Única/métodos , Resposta a Proteínas não Dobradas/genética
7.
Nat Biotechnol ; 37(7): 810-818, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31267104

RESUMO

A major challenge for stem cell engineering is achieving a holistic understanding of the molecular networks and biological processes governing cell differentiation. To address this challenge, we describe a computational approach that combines gene expression analysis, previous knowledge from proteomic pathway informatics and cell signaling models to delineate key transitional states of differentiating cells at high resolution. Our network models connect sparse gene signatures with corresponding, yet disparate, biological processes to uncover molecular mechanisms governing cell fate transitions. This approach builds on our earlier CellNet and recent trajectory-defining algorithms, as illustrated by our analysis of hematopoietic specification along the erythroid lineage, which reveals a role for the EGF receptor family member, ErbB4, as an important mediator of blood development. We experimentally validate this prediction and perturb the pathway to improve erythroid maturation from human pluripotent stem cells. These results exploit an integrative systems perspective to identify new regulatory processes and nodes useful in cell engineering.


Assuntos
Engenharia Celular , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Pluripotentes Induzidas/fisiologia , Biologia de Sistemas/métodos , Algoritmos , Animais , Antígenos CD34/genética , Antígenos CD34/metabolismo , Diferenciação Celular , Linhagem da Célula , Proliferação de Células , Biologia Computacional/métodos , Eritrócitos , Eritropoese , Citometria de Fluxo , Regulação da Expressão Gênica , Redes Reguladoras de Genes , Humanos , Camundongos , Receptor ErbB-4/metabolismo , Transdução de Sinais , Peixe-Zebra
8.
Cancer Invest ; 37(6): 242-252, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31296070

RESUMO

Drug resistance to TKIs and the existance of CML leukemia stem cells is an urgent problem. In this study, we demonstrate that quinacrine (QC) induces apoptosis in BCR-ABL positive CML and acute lymphoblastic leukemia (ALL) cells. Interestingly, QC inhibits the colony formation of primary CD34+ progenitor/stem leukemia cells from CML patients. QC targets RNA polymerase I, which produces ribosomal (r)RNA, involving in protein translation process. Also, QC treatment prolongs CML-like mice survival and inhibits K562 tumor growth in vivo. In conclusion, we demonstrate that QC depletes BCR-ABL protein and suppresses Ph-positive leukemia cells in vitro and in vivo.


Assuntos
Proteínas de Fusão bcr-abl/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Quinacrina/uso terapêutico , Animais , Antígenos CD34/metabolismo , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Feminino , Humanos , Células K562 , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos NOD , Camundongos SCID , Células-Tronco Neoplásicas/efeitos dos fármacos , Células-Tronco Neoplásicas/metabolismo , Inibidores de Proteínas Quinases/uso terapêutico
9.
Nat Commun ; 10(1): 2395, 2019 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-31160568

RESUMO

Hematopoietic Stem/Progenitor cells (HSPCs) are endowed with the role of maintaining a diverse pool of blood cells throughout the human life. Despite recent efforts, the nature of the early cell fate decisions remains contentious. Using single-cell RNA-Seq, we show that existing approaches to stratify bone marrow CD34+ cells reveal a hierarchically-structured transcriptional landscape of hematopoietic differentiation. Still, this landscape misses important early fate decisions. We here provide a broader transcriptional profiling of bone marrow lineage negative hematopoietic progenitors that recovers a key missing branchpoint into basophils and expands our understanding of the underlying structure of early adult human haematopoiesis. We also show that this map has strong similarities in topology and gene expression to that found in mouse. Finally, we identify the sialomucin CD164, as a reliable marker for the earliest branches of HSPCs specification and we showed how its use can foster the design of alternative transplantation cell products.


Assuntos
Hematopoese/genética , Células-Tronco Hematopoéticas/metabolismo , Animais , Antígenos CD34/metabolismo , Células da Medula Óssea , Linhagem da Célula , Endolina/metabolismo , Perfilação da Expressão Gênica , Humanos , Camundongos , Análise de Sequência de RNA , Análise de Célula Única
11.
Anticancer Res ; 39(6): 2739-2747, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31177109

RESUMO

BACKGROUND/AIM: The aim of the present study was to investigate the vascular normalization effect of traditional Chinese medicine Astragalus membranaceus (AM) and Curcuma wenyujin (CW) on tumor-derived endothelial cells (TECs). MATERIALS AND METHODS: TECs were isolated from the xenografted HCC cell line HepG2 expressing red fluorescent protein (RFP). The effect of AM and CW on TECs proliferation was measured using the CCK8 assay. The vascular normalization potential of AM and CW was assessed using a tube formation assay. Immunocytochemistry was performed to assess the effect of AM and CW on the expression of angiogenic maker CD34 and hypoxia-inducible factor HIF1a. RESULTS: The isolated TECs and endothelioma (EOMA) cells did not differ with regard to the expression levels of endothelial markers CD34, VEGFR-1, VEGFR-2, PDGFR-α and PDGFR-ß. All AM, CW, AM+CW and Nintedanib (Nin) showed a dose-dependent increasing inhibition effect on either TECs or EOMA cells. AM, CW and AM+CW significantly reduced HIF1a expression, increased CD34 expression and enhanced endothelial network formation in TECs or EOMA cells compared to the control. CONCLUSION: AM and CW promoted vascular normalization in tumor-derived endothelial cells of HCC, through increased expression of CD34 and reduced expression of HIF1a.


Assuntos
Antígenos CD34/metabolismo , Carcinoma Hepatocelular/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Células Endoteliais/citologia , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Neoplasias Hepáticas/metabolismo , Animais , Astragalus propinquus/química , Carcinoma Hepatocelular/irrigação sanguínea , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Curcuma/química , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/irrigação sanguínea , Medicina Tradicional Chinesa , Camundongos , Transplante de Neoplasias , Transdução de Sinais/efeitos dos fármacos
12.
Life Sci ; 231: 116521, 2019 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-31152814

RESUMO

INTRODUCTION: Telocytes (TCs) are recently described to integrate a variety of different cells. AIM OF THE WORK: The aim was to investigate the presence of TCs in the rat mammary gland at its different physiological stages. MATERIAL AND METHODS: Twenty four adult female albino rats were classified into 4 groups: resting, mid-pregnancy, lactating, and involution groups. Inguinal mammary glands were processed for immunohistochemical and transmission electron microscopic (TEM) examination. RESULTS: TCs were immune-positive for c-kit and CD34 and showed significant differences in the different studied groups indicating variable roles at the different stages. TEM results characterized TCs by its shape and the long slender and moniliform telopodes linking the cells into stromal networks. The extracellular exosomes, homo-cellular synapsis and hetero-cellular synapsis were observed. CONCLUSION: Our study provides evidence for the presence of TCs in all stages of the gland; not only in the resting stage as proved by other studies, but with immune-labeling differences suggesting different structural and physiological roles of TCs according to the stage requirements. These functions might via controlling the proliferation during pregnancy and lactation and the involution of the gland after weaning. Thus, more future functional studies of TCs will be important to help understanding the mechanism by which TCs contribute to tissue homeostasis concerning the role of the stromal/epithelial interactions in mammary gland biology and pathology including breast cancer which would be revolutionary for future therapeutic applications.


Assuntos
Glândulas Mamárias Animais/fisiologia , Telócitos/fisiologia , Telócitos/ultraestrutura , Animais , Antígenos CD34/metabolismo , Antígenos CD34/fisiologia , Tecido Conjuntivo , Feminino , Imuno-Histoquímica , Lactação , Glândulas Mamárias Animais/citologia , Microscopia Eletrônica de Transmissão , Gravidez , Proteínas Proto-Oncogênicas c-kit/metabolismo , Proteínas Proto-Oncogênicas c-kit/fisiologia , Ratos , Ratos Wistar
13.
J Med Life ; 12(1): 30-33, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31123522

RESUMO

Glomus tumors are frequently associated with pain, tenderness and cold sensitivity. We report the diagnosis and successful surgical management of a case of a classic glomus tumor in a young woman. The clinical diagnosis was made on the basis of medical history and MRI findings. The lesion was excised via a dorsolateral subungual approach, leading to the complete resolution of symptoms. Histology confirmed the lesion to be a glomus tumor. Glomus tumors are painful subungual lesions. They produce a throbbing or lancinating local discomfort, cold-sensitivity, and severe pain following minor trauma. The diagnosis is confirmed by histology, but the clinical diagnosis is highly suggestive. Complete excision will usually relieve pain. Recurrence is common following incomplete resection.


Assuntos
Dedos/patologia , Tumor Glômico/patologia , Dor/etiologia , Adulto , Antígenos CD34/metabolismo , Proliferação de Células , Feminino , Tumor Glômico/diagnóstico por imagem , Tumor Glômico/cirurgia , Humanos , Antígeno Ki-67/metabolismo , Imagem por Ressonância Magnética , Mitose , Polegar/diagnóstico por imagem , Polegar/cirurgia
14.
Breast Cancer Res Treat ; 176(2): 387-394, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31041685

RESUMO

PURPOSE: Circulating adipose stromal cells (CASC) are thought to be increased in obesity and facilitate angiogenesis, and tumor metastases. METHODS: CASC were identified from buffy coat peripheral blood mononuclear cells (PBMCs) by flow cytometry as CD34brightCD31- CD45- and CASC frequency was compared to adiposity measures in 33 women at increased risk for breast cancer. Feasibility of CASC as a response biomarker for a diet and exercise intervention in ten breast cancer survivors was then explored. RESULTS: For 33 high-risk women, median CASC frequency was 9.7 per million PBMCs and trended positively with body mass index, fat mass index (FMI), and percent android fat. Correlation was significant when BMI was dichotomized at > versus < 35 kg/m2 (p = 0.02). For ten breast cancer survivors with a median BMI of 37 kg/m2, median CASC frequency was 16.4 per million PBMCs. In univariate analyses, change in BMI, total fat and visceral fat were significantly correlated with change in CASC frequency. On multivariate analysis, change in visceral adipose had the strongest association with change in CASC frequency (p < 0.00078). CONCLUSIONS: The association between the reduction in visceral adipose tissue and the decrease in frequency of circulating adipose stromal cells suggests that the latter might be a useful biomarker in clinical trials of obese breast cancer survivors undergoing a weight loss intervention.


Assuntos
Tecido Adiposo/imunologia , Biomarcadores/sangue , Neoplasias da Mama/sangue , Obesidade/terapia , Tecido Adiposo/citologia , Idoso , Antígenos CD34/metabolismo , Neoplasias da Mama/imunologia , Sobreviventes de Câncer , Estudos Transversais , Dietoterapia , Terapia por Exercício , Feminino , Humanos , Antígenos Comuns de Leucócito/metabolismo , Pessoa de Meia-Idade , Obesidade/sangue , Obesidade/imunologia , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Pós-Menopausa , Pré-Menopausa , Células Estromais/citologia , Células Estromais/imunologia
15.
J Nanobiotechnology ; 17(1): 62, 2019 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-31084605

RESUMO

BACKGROUND: Human hematopoietic progenitor cells (HPCs) are important for cell therapy in cancer and tissue regeneration. In vitro studies have shown a transient association of 40 nm polystyrene nanoparticles (PS NPs) with these cells, which is of interest for intelligent design and application of NPs in HPC-based regenerative protocols. In this study, we aimed to investigate the involvement of nanoparticles' size and membrane-attached glycan molecules in the interaction of HPCs with PS NPs, and compared it with monocytes. Human cord blood-derived HPCs and THP-1 cells were exposed to fluorescently labelled, carboxylated PS NPs of 40, 100 and 200 nm. Time-dependent nanoparticle membrane association and/or uptake was observed by measuring fluorescence intensity of exposed cells at short time intervals using flow cytometry. By pretreating the cells with neuraminidase, we studied the possible effect of membrane-associated sialic acids in the interaction with NPs. Confocal microscopy was used to visualize the cell-specific character of the NP association. RESULTS: Confocal images revealed that the majority of PS NPs was initially observed to be retained at the outer membrane of HPCs, while the same NPs showed immediate internalization by THP-1 monocytic cells. After prolonged exposure up to 4 h, PS NPs were also observed to enter the HPCs' intracellular compartment. Cell-specific time courses of NP association with HPCs and THP-1 cells remained persistent after cells were enzymatically treated with neuraminidase, but significantly increased levels of NP association could be observed, suggesting a role for membrane-associated sialic acids in this process. CONCLUSIONS: We conclude that the terminal membrane-associated sialic acids contribute to the NP retention at the outer cell membrane of HPCs. This retention behavior is a unique characteristic of the HPCs and is independent of NP size.


Assuntos
Células-Tronco Hematopoéticas/metabolismo , Monócitos/metabolismo , Nanopartículas/química , Ácidos Siálicos/química , Antígenos CD34/metabolismo , Transporte Biológico , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Preparações de Ação Retardada/metabolismo , Endocitose/efeitos dos fármacos , Humanos , Tamanho da Partícula , Poliestirenos , Propriedades de Superfície
16.
Nat Commun ; 10(1): 1634, 2019 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-30967552

RESUMO

Gene correction in human long-term hematopoietic stem cells (LT-HSCs) could be an effective therapy for monogenic diseases of the blood and immune system. Here we describe an approach for X-linked sSevere cCombined iImmunodeficiency (SCID-X1) using targeted integration of a cDNA into the endogenous start codon to functionally correct disease-causing mutations throughout the gene. Using a CRISPR-Cas9/AAV6 based strategy, we achieve up to 20% targeted integration frequencies in LT-HSCs. As measures of the lack of toxicity we observe no evidence of abnormal hematopoiesis following transplantation and no evidence of off-target mutations using a high-fidelity Cas9 as a ribonucleoprotein complex. We achieve high levels of targeting frequencies (median 45%) in CD34+ HSPCs from six SCID-X1 patients and demonstrate rescue of lymphopoietic defect in a patient derived HSPC population in vitro and in vivo. In sum, our study provides specificity, toxicity and efficacy data supportive of clinical development of genome editing to treat SCID-Xl.


Assuntos
DNA Complementar/genética , Edição de Genes/métodos , Transplante de Células-Tronco Hematopoéticas , Subunidade gama Comum de Receptores de Interleucina/genética , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/terapia , Animais , Antígenos CD34/metabolismo , Sistemas CRISPR-Cas/genética , Linhagem Celular , Códon de Iniciação/genética , Éxons/genética , Sangue Fetal/citologia , Vetores Genéticos/genética , Voluntários Saudáveis , Células-Tronco Hematopoéticas/metabolismo , Humanos , Masculino , Camundongos , Mutação , Parvovirinae/genética , Cultura Primária de Células , Fatores de Tempo , Transdução Genética/métodos , Quimeras de Transplante/genética , Transplante Heterólogo/métodos , Doenças por Imunodeficiência Combinada Ligada ao Cromossomo X/genética
17.
Anal Cell Pathol (Amst) ; 2019: 5134156, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30963020

RESUMO

Aim: This study is aimed at investigating the effect of Shogaol, a phenolic constituent of ginger, on dextran sodium sulfate- (DSS-) induced ulcerative colitis (UC) in mice in comparison with 6-thioguanine (6-TG), an immune-suppressant chemotherapeutic medicine used for treatment of ulcerative colitis. Material & Methods: Thirty-six adult, male and female BALB/c mice were randomly divided into six groups: group 1 (control negative) not exposed to DSS and did not receive any treatment, group 2 (control positive) exposed to DSS but did not receive any treatment, group 3 exposed to DSS and treated by 0.1 mg/kg of 6-thioguanine, and groups 4, 5, and 6 exposed to DSS and treated by 10, 20, and 40 mg/kg b.w. Shogaol, respectively. At day 56, the mice were checked for their disease activity index (DAI) and they were sacrificed. The colons of the mice were examined for length measurement, histological index score, and the expression of CD133 and CD34 stem cell markers. Results: Shogaol showed a better curative effect than did 6-TG in repairing the colonic mucosal damages in DSS-exposed mice as indicated by the levels of CD133 and CD34 expression in the colonic crypts and by the DAI score, colon length measurements, & histological index score which were significantly reduced in mice treated by Shogaol, particularly the 20 and 40 mg/kg BW doses. Conclusion: The results of this study indicated that oral treatment with the ginger-derived substance Shogaol could be better than the conventional immunosuppressive chemotherapeutic remedy 6-TG in treatment of DSS-induced UC.


Assuntos
Catecóis/uso terapêutico , Colite/tratamento farmacológico , Colite/metabolismo , Mucosa Intestinal/citologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Antígeno AC133/metabolismo , Animais , Antígenos CD34/metabolismo , Feminino , Imuno-Histoquímica , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Análise Serial de Tecidos
18.
PLoS Genet ; 15(4): e1008034, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-31017901

RESUMO

Melanocyte stem cells (McSCs) are the undifferentiated melanocytic cells of the mammalian hair follicle (HF) responsible for recurrent generation of a large number of differentiated melanocytes during each HF cycle. HF McSCs reside in both the CD34+ bulge/lower permanent portion (LPP) and the CD34- secondary hair germ (SHG) regions of the HF during telogen. Using Dct-H2BGFP mice, we separate bulge/LPP and SHG McSCs using FACS with GFP and anti-CD34 to show that these two subsets of McSCs are functionally distinct. Genome-wide expression profiling results support the distinct nature of these populations, with CD34- McSCs exhibiting higher expression of melanocyte differentiation genes and with CD34+ McSCs demonstrating a profile more consistent with a neural crest stem cell. In culture and in vivo, CD34- McSCs regenerate pigmentation more efficiently whereas CD34+ McSCs selectively exhibit the ability to myelinate neurons. CD34+ McSCs, and their counterparts in human skin, may be useful for myelinating neurons in vivo, leading to new therapeutic opportunities for demyelinating diseases and traumatic nerve injury.


Assuntos
Antígenos CD34/metabolismo , Melanócitos/imunologia , Melanócitos/fisiologia , Células-Tronco/imunologia , Células-Tronco/fisiologia , Animais , Diferenciação Celular , Células Cultivadas , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Cor de Cabelo/fisiologia , Folículo Piloso/citologia , Folículo Piloso/fisiologia , Melanócitos/classificação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Camundongos Nus , Camundongos Transgênicos , Proteína Básica da Mielina/deficiência , Proteína Básica da Mielina/genética , Crista Neural/citologia , Crista Neural/imunologia , Crista Neural/fisiologia , Pigmentação/fisiologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Regeneração/fisiologia , Células-Tronco/classificação
19.
EBioMedicine ; 43: 150-158, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30975542

RESUMO

BACKGROUND: Systemic mastocytosis (SM) is a haematological disease characterised by organ infiltration by neoplastic mast cells. Almost all SM patients have a mutation in the gene encoding the tyrosine kinase receptor KIT causing a D816V substitution and autoactivation of the receptor. Mast cells and CD34+ haematopoietic progenitors can carry the mutation; however, in which progenitor cell subset the mutation arises is unknown. We aimed to investigate the distribution of the D816V mutation in single mast cells and single haematopoietic stem and progenitor cells. METHODS: Fluorescence-activated single-cell index sorting and KIT D816V mutation assessment were applied to analyse mast cells and >10,000 CD34+ bone marrow progenitors across 10 haematopoietic progenitor subsets. In vitro assays verified cell-forming potential. FINDINGS: We found that in SM 60-99% of the mast cells harboured the KIT D816V mutation. Despite increased frequencies of mast cells in SM patients compared with control subjects, the haematopoietic progenitor subset frequencies were comparable. Nevertheless, the mutation could be detected throughout the haematopoietic landscape of SM patients, from haematopoietic stem cells to more lineage-primed progenitors. In addition, we demonstrate that FcεRI+ bone marrow progenitors exhibit mast cell-forming potential, and we describe aberrant CD45RA expression on SM mast cells for the first time. INTERPRETATION: The KIT D816V mutation arises in early haematopoietic stem and progenitor cells and the mutation frequency is approaching 100% in mature mast cells, which express the aberrant marker CD45RA.


Assuntos
Substituição de Aminoácidos , Predisposição Genética para Doença , Células-Tronco Hematopoéticas/metabolismo , Mastocitose Sistêmica/etiologia , Mutação , Proteínas Proto-Oncogênicas c-kit/genética , Antígenos CD34/metabolismo , Biomarcadores , Células da Medula Óssea/metabolismo , Células Cultivadas , Ensaio de Unidades Formadoras de Colônias , Citometria de Fluxo , Estudos de Associação Genética , Humanos , Imunofenotipagem , Antígenos Comuns de Leucócito/metabolismo , Mastócitos/imunologia , Mastócitos/metabolismo , Mastocitose Sistêmica/diagnóstico , Mastocitose Sistêmica/metabolismo , Análise de Célula Única
20.
Chem Biol Interact ; 305: 40-47, 2019 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-30894315

RESUMO

Diabetes mellitus and periodontitis have long been considered to be biologically linked. Erythropoietin (EPO) has multiple biological functions, such as stimulating the proliferation and differentiation of erythroid progenitor cells and reducing glucose-induced oxidative stress via different mechanisms, acting as a direct antioxidant. The purposes of the study to examine the anti-oxidative effect of EPO on reducing high glucose-induced oxidative stress of hPDLSCs and provide a better understanding of the mechanism of these processes. PDLSCs were induced by highglucose (HG, 30 mM) in the presence or absence of EPO. Cell proliferation was measured by MTT assay. The reactiveoxygen species (ROS) level, malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were detected to evaluate oxidative stress. qRT-PCR and western blot analysis were used to examine the expression of osteogenic related genes and protein (Runx2 and Osterix). Alizarin Red-S staining was used to detect mineralized nodule formation. The results showed that EPO promote the proliferation of PDLSCs, which was suppressed by high glucose (30 mM). Moreover, EPO attenuated high glucose (30 mM) induced oxidative stress by reducing the levels of ROS and MDA, and increasing the SOD activity. Furthermore,EPO alleviate high glucose(30 mM) induced suppression of osteogenic differentiation ability in PDLSCs, as evidenced by the up-regulated mRNA and protein expression of Runx2 and Osterix and increased ALP activity. In conclusion, EPO attenuates high glucose-induced oxidative stress, inhibitory effect of proliferation and inhibition of osteogenic differentiation in periodontal ligament stem cell (PDLSCs).


Assuntos
Diferenciação Celular/efeitos dos fármacos , Eritropoetina/farmacologia , Glucose/farmacologia , Osteogênese/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Adolescente , Antígenos CD34/metabolismo , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Humanos , Malondialdeído/metabolismo , Ligamento Periodontal/citologia , Espécies Reativas de Oxigênio/metabolismo , Fator de Transcrição Sp7/genética , Fator de Transcrição Sp7/metabolismo , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Superóxido Dismutase/metabolismo , Adulto Jovem
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