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1.
Clin Chim Acta ; 495: 538-544, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31145896

RESUMO

CXC chemokine ligand 12 (CXCL12) is a specific chemokine ligand and plays a significant role in cell chemotaxis. Upon binding to CXC chemokine receptor 4 (CXCR4) or CXCR7, CXCL12 can activate different signaling cascades to regulate cell proliferation, migration, and metabolism. CXCL12 exerts a pro-atherogenic action by aggravating multiple pathogenesis of atherogenesis, including dyslipidemia, inflammation, neointima hyperplasia, angiogenesis, and insulin resistance. Serum CXCL12 levels are also markedly increased in patients with atherosclerosis-associated disease. The present review focuses on recent advances in CXCL12 research in the pathogenesis of atherosclerosis together with its clinical values. This may provide insight into potential novel therapies for atherosclerosis.


Assuntos
Aterosclerose/tratamento farmacológico , Aterosclerose/metabolismo , Quimiocina CXCL12/metabolismo , Terapia de Alvo Molecular/métodos , Animais , Aterosclerose/patologia , Quimiocina CXCL12/biossíntese , Humanos , Transdução de Sinais/efeitos dos fármacos
3.
Vasc Med ; 24(3): 200-207, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30786835

RESUMO

The efficacy of biologic therapies in critical limb ischemia (CLI) remains elusive, in part, due to limitations in trial design and patient selection. Using a novel design, we examined the impact of complementing revascularization therapy with intramuscular JVS-100 - a non-viral gene therapy that activates endogenous regenerative repair pathways. In this double-blind, placebo-controlled, Phase 2B trial, we randomized 109 patients with CLI (Rutherford class V or VI) to 8 mg or 16 mg intramuscular injections of placebo versus JVS-100. Patients were eligible if they persistently had reduced forefoot perfusion, by toe-brachial index (TBI) or skin perfusion pressure (SPP), following successful revascularization with angiographic demonstration of tibial arterial flow to the ankle. The primary efficacy end point was a 3-month wound healing score assessed by an independent wound core laboratory. The primary safety end point was major adverse limb events (MALE). Patients' mean age was 71 years, 33% were women, 79% had diabetes, and 8% had end-stage renal disease. TBI after revascularization was 0.26, 0.27, and 0.26 among the three groups (placebo, 8 mg, and 16 mg injections, respectively). Only 26% of wounds completely healed at 3 months, without any differences between the three groups (26.5%, 26.5%, and 25%, respectively). Similarly, there were no significant changes in TBI at 3 months. Three (2.8%) patients died and two (1.8%) had major amputations. Rates of MALE at 3 months were 8.8%, 20%, and 8.3%, respectively. While safe, JVS-100 failed to improve wound healing or hemodynamic measures at 3 months. Only one-quarter of CLI wounds healed at 3 months despite successful revascularization, highlighting the need for additional research in therapies that can improve microcirculation in these patients. ClinicalTrials.gov Identifier: NCT02544204.


Assuntos
Quimiocina CXCL12/genética , Terapia Genética/métodos , Hemodinâmica , Doença Arterial Periférica/terapia , Plasmídeos/genética , Idoso , Idoso de 80 Anos ou mais , Índice Tornozelo-Braço , Quimiocina CXCL12/biossíntese , Método Duplo-Cego , Feminino , Humanos , Masculino , Microcirculação , Pessoa de Meia-Idade , Doença Arterial Periférica/diagnóstico , Doença Arterial Periférica/genética , Doença Arterial Periférica/metabolismo , Fluxo Sanguíneo Regional , Fatores de Tempo , Resultado do Tratamento , Estados Unidos , Cicatrização
4.
Prostate ; 79(7): 757-767, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30811623

RESUMO

BACKGROUND: Several studies show that prostatic fibrosis is associated with male lower urinary tract dysfunction (LUTD). Development of fibrosis is typically attributed to signaling through the transforming growth factor ß (TGF-ß) pathway, but our laboratory has demonstrated that in vitro treatment of human prostatic fibroblasts with the C-X-C motif chemokine ligand 12 (CXCL12) chemokine stimulates myofibroblast phenoconversion and that CXCL12 has the capacity to activate profibrotic pathways in these cells in a TGF-ß-independent manner. We have previously reported that feeding mice high-fat diet (HFD) results in obesity, type II diabetes, increased prostatic fibrosis, and urinary voiding dysfunction. The purpose of this study was to test the hypothesis that in vivo blockade of the CXCL12/CXCR4 axis would inhibit the development of fibrosis-mediated LUTD in HFD-fed mice. METHODS: Two-month-old male senescence-accelerated mouse prone-6 mice were fed either a HFD or low-fat diet (LFD) for 8 months. Half of each dietary group were given constant access to normal water or water that contained the C-X-C chemokine receptor type 4 (CXCR4; CXCL12 receptor) antagonist CXCR4AIII. At the conclusion of the study, mice were weighed, subjected to oral glucose tolerance testing and cystometry, and lower urinary tract tissues collected and assessed for collagen content. RESULTS: HFD-fed mice became significantly obese, insulin resistant, and hyperglycemic, consistent with acquisition of metabolic syndrome, compared with LFD-fed mice. Anesthetized cystometry demonstrated that HFD-fed mice experienced significantly longer intercontractile intervals and greater functional bladder capacity than LFD-fed mice. Immunohistochemistry demonstrated high levels of CXCR4 and CXCR7 staining in mouse prostate epithelial and stromal cells. Picrosirius red staining indicated significantly greater periurethral collagen deposition in the prostates of HFD than LFD-fed mice. Treatment with the CXCR4 antagonist CXCR4AIII did not affect acquisition of metabolic syndrome but did reduce both urinary voiding dysfunction and periurethral prostate collagen accumulation. CONCLUSIONS: This is the first study to report that obesity-induced lower urinary tract fibrosis and voiding dysfunction can be repressed by antagonizing the activity of the CXCR4 chemokine receptor in vivo. These data suggest that targeting the CXCL12/CXCR4 signaling pathway may be a clinical option for the prevention or treatment of human male LUTD.


Assuntos
Anti-Inflamatórios/farmacologia , Sintomas do Trato Urinário Inferior/tratamento farmacológico , Próstata/efeitos dos fármacos , Receptores CXCR4/antagonistas & inibidores , Receptores CXCR4/biossíntese , Animais , Quimiocina CXCL12/antagonistas & inibidores , Quimiocina CXCL12/biossíntese , Colágeno/metabolismo , Dieta Hiperlipídica/efeitos adversos , Modelos Animais de Doenças , Fibrose/etiologia , Fibrose/patologia , Sintomas do Trato Urinário Inferior/etiologia , Sintomas do Trato Urinário Inferior/fisiopatologia , Masculino , Síndrome Metabólica/etiologia , Camundongos , Obesidade/etiologia , Próstata/patologia , Transdução de Sinais/efeitos dos fármacos
5.
Neurosci Res ; 144: 48-55, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30120960

RESUMO

Chemokines related neuroinflammation and N-methyl-d-aspartate receptor (NMDAR) mediated nociceptive transmission are pivotal determinants in the pathogenesis of opioid-induced hyperalgesia (OIH), but little is known about specific mechanism and treatment. Chemokine CXCL12 with its receptor CXCR4 is implicated in different pathological pain, moreover, neurotoxicity of CXCL12 is associated with NMDAR activation. Recent studies recapitulate the anti-nociception of Annexin 1 (ANXA1) in inflammatory pain. This study examined whether ANXA1 prevented remifentanil-caused OIH through modulating CXCL12 and NMDAR pathway in rats. Acute exposure to remifentanil induced mechanical allodynia and thermal hyperalgesia, which was accompanied by the increase of spinal ANXA1 and CXCL12/CXCR4 expression. Central injection of Anxa12-26 attenuated behavioral OIH in a dose-dependent manner, facilitated ANXA1 production, and inhibited up-regulation of CXCL12/CXCR4 level and NR2B-containing NMDAR phosphorylation. Moreover, pretreatment with AMD3100 reduced hyperalgesia and NR2B-containing NMDAR phosphorylation. Also, exogenous CXCL12 elicited pain hypersensitivity and NMDAR activation in naïve rats, which was reversed by the supplemental delivery of Anxa12-26. These current findings indicate the participation of spinal CXCL12/CXCR4 and NR2B-containing NMDAR pathway in anti-hyperalgesic action of ANXA1 in OIH.


Assuntos
Anexina A1/farmacologia , Quimiocina CXCL12/metabolismo , Receptores CXCR4/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Remifentanil/antagonistas & inibidores , Analgésicos Opioides/antagonistas & inibidores , Analgésicos Opioides/farmacologia , Animais , Anexina A1/metabolismo , Quimiocina CXCL12/antagonistas & inibidores , Quimiocina CXCL12/biossíntese , Interações de Medicamentos , Compostos Heterocíclicos/farmacologia , Hiperalgesia/induzido quimicamente , Hiperalgesia/tratamento farmacológico , Hiperalgesia/metabolismo , Masculino , Fragmentos de Peptídeos/farmacologia , Fosforilação/efeitos dos fármacos , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Receptores CXCR4/antagonistas & inibidores , Receptores CXCR4/biossíntese , Remifentanil/farmacologia , Transdução de Sinais/efeitos dos fármacos
6.
Sci Rep ; 8(1): 14501, 2018 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-30266921

RESUMO

Skeletal osteoblasts are important regulators of B-lymphopoiesis, serving as a rich source of factors such as CXCL12 and IL-7 which are crucial for B-cell development. Recent studies from our laboratory and others have shown that deletion of Rptor, a unique component of the mTORC1 nutrient-sensing complex, early in the osteoblast lineage development results in defective bone development in mice. In this study, we now demonstrate that mTORC1 signalling in pre-osteoblasts is required for normal B-lymphocyte development in mice. Targeted deletion of Rptor in osterix-expressing pre-osteoblasts (Rptorob-/-) leads to a significant reduction in the number of B-cells in the bone marrow, peripheral blood and spleen at 4 and 12 weeks of age. Rptorob-/- mice also exhibit a significant reduction in pre-B and immature B-cells in the BM, indicative of a block in B-cell development from the pro-B to pre-B cell stage. Circulating levels of IL-7 and CXCL12 are also significantly reduced in Rptorob-/- mice. Importantly, whilst Rptor-deficient osteoblasts are unable to support HSC differentiation to B-cells in co-culture, this can be rescued by the addition of exogenous IL-7 and CXCL12. Collectively, these findings demonstrate that mTORC1 plays an important role in extrinsic osteoblastic regulation of B-cell development.


Assuntos
Linfócitos B/citologia , Linfopoese/fisiologia , Alvo Mecanístico do Complexo 1 de Rapamicina/fisiologia , Osteoblastos/metabolismo , Animais , Linfócitos B/metabolismo , Quimiocina CXCL12/biossíntese , Quimiocina CXCL12/sangue , Quimiocina CXCL12/farmacologia , Técnicas de Cocultura , Regulação para Baixo , Genes Reporter , Interleucina-7/sangue , Interleucina-7/farmacologia , Camundongos , Camundongos Knockout , Camundongos Transgênicos , RNA Mensageiro/biossíntese , Proteína Regulatória Associada a mTOR/deficiência , Proteína Regulatória Associada a mTOR/genética , Proteína Regulatória Associada a mTOR/fisiologia , Fator de Transcrição Sp7/metabolismo
7.
Artigo em Inglês | MEDLINE | ID: mdl-30096932

RESUMO

The effect of horticultural therapy (HT) on immune and endocrine biomarkers remains largely unknown. We designed a waitlist-control randomized controlled trial to investigate the effectiveness of HT in improving mental well-being and modulating biomarker levels. A total of 59 older adults was recruited, with 29 randomly assigned to the HT intervention and 30 to the waitlist control group. The participants attended weekly intervention sessions for the first 3 months and monthly sessions for the subsequent 3 months. Biological and psychosocial data were collected. Biomarkers included IL-1ß, IL-6, sgp-130, CXCL12/SDF-1α, CCL-5/RANTES, BDNF (brain-derived neurotrophic factor), hs-CRP, cortisol and DHEA (dehydroepiandrosterone). Psychosocial measures examined cognitive functions, depression, anxiety, psychological well-being, social connectedness and satisfaction with life. A significant reduction in plasma IL-6 level (p = 0.02) was observed in the HT intervention group. For the waitlist control group, significant reductions in plasma CXCL12 (SDF-1α) (p = 0.003), CXCL5 (RANTES) (p = 0.05) and BDNF (p = 0.003) were observed. A significant improvement in social connectedness was also observed in the HT group (p = 0.01). CONCLUSION: HT, in reducing plasma IL-6, may prevent inflammatory disorders and through maintaining plasma CXCL12 (SDF-1α), may maintain hematopoietic support to the brain. HT may be applied in communal gardening to enhance the well-being of older adults.


Assuntos
Grupo com Ancestrais do Continente Asiático , Horticultura Terapêutica/métodos , Saúde Mental , Idoso , Ansiedade/epidemiologia , Encéfalo/metabolismo , Fator Neurotrófico Derivado do Encéfalo/biossíntese , Quimiocina CXCL12/biossíntese , Cognição , Desidroepiandrosterona/biossíntese , Depressão/epidemiologia , Feminino , Humanos , Hidrocortisona/biossíntese , Interleucinas/biossíntese , Relações Interpessoais , Masculino , Pessoa de Meia-Idade
8.
Exp Neurol ; 306: 55-63, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29715476

RESUMO

Addiction and rewarding effect is a primary side effect of morphine, which is commonly used to relieve the acute or chronic pain. Several lines of evidence have suggested that inflammation response in the VTA contributes to morphine-induced reward (conditioned place preference, CPP), while the mechanism are poorly understood. The present study showed that repeated morphine conditioning persistently increased the expression of CXCL12 mRNA and protein in VTA. Furthermore, inhibition of CXCL12 prevented the acquisition and maintenance, but not the expression, of morphine-induced CPP in rodent. In addition, molecular analysis revealed that morphine conditioning increased the occupancy of p-STAT3 in the specific binding site (-1667/-1685) of CXCL12 promoter regions, and enhanced the interaction between acetyltransferase p300 and STAT3, and, hence, induced the histone H4 hyperacetylation in the promoter region and facilitated the transcription and expression of CXCL12 in VTA. Collectively, these results, for the first time, provided the evidence that persisted increase of VTA CXCL12 via epigenetic mechanism mediated the acquisition and maintenance, but not the expression, of morphine CPP.


Assuntos
Quimiocina CXCL12/genética , Condicionamento Operante/efeitos dos fármacos , Epigênese Genética/genética , Morfina/farmacologia , Entorpecentes/farmacologia , Animais , Quimiocina CXCL12/biossíntese , Regulação da Expressão Gênica , Histonas/metabolismo , Imuno-Histoquímica , Masculino , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Interferente Pequeno/farmacologia , Ratos , Ratos Sprague-Dawley , Fator de Transcrição STAT3/biossíntese , Fator de Transcrição STAT3/genética , Regulação para Cima
9.
Exp Hematol ; 64: 59-70.e2, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29775645

RESUMO

Mesenchymal stromal cells (MSCs) support hematopoietic stem cells (HSCs) in vivo and enhance HSC engraftment and hematopoietic recovery upon cotransplantation with HSCs. These data have led to the hypothesis that MSCs may affect the HSC niche, leading to changes in HSC retention and trafficking. We studied the effect of MSC administration on the HSC compartment in the bone marrow (BM) in mice. After injection of MSCs, HSC numbers in the BM were decreased coinciding with an increased cell cycle activity compared with phosphate-buffered saline (PBS)-injected controls. Furthermore, the frequency of macrophages was significantly reduced and niche factors including Cxcl12, Scf, and Vcam were downregulated in endosteal cells. These BM changes are reminiscent of events associated with granulocyte colony-stimulating factor (G-CSF)-induced hematopoietic stem and progenitor cell (HSPC) mobilization. Interestingly, coadministration of MSCs and G-CSF resulted in a twofold increase in peripheral blood HSPC release compared with injection of G-CSF alone, whereas injection of MSCs alone did not induce HSPC mobilization. After intravenous administration, MSCs were only observed in the lungs, suggesting that they exert their effect on the HSC niche through a soluble mediator. Therefore, we tested the hypothesis that MSC-derived extracellular vesicles (EVs) are responsible for the observed changes in the HSC niche. Indeed, administration of EVs resulted in downregulation of Cxcl12, Scf, and Vcam and enhanced G-CSF-induced HSPC mobilization at similar levels as MSCs and G-CSF. Together, these data indicate that MSCs induce a permissive state in the BM, enhancing HSPC mobilization through the release of EVs.


Assuntos
Medula Óssea/fisiologia , Vesículas Extracelulares/fisiologia , Fator Estimulador de Colônias de Granulócitos/farmacologia , Mobilização de Células-Tronco Hematopoéticas/métodos , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/fisiologia , Animais , Quimiocina CXCL12/biossíntese , Quimiocina CXCL12/genética , Citocinas/farmacologia , Regulação da Expressão Gênica , Humanos , Macrófagos/citologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Recombinantes/farmacologia , Fator de Células-Tronco/biossíntese , Fator de Células-Tronco/genética , Nicho de Células-Tronco , Molécula 1 de Adesão de Célula Vascular/biossíntese , Molécula 1 de Adesão de Célula Vascular/genética
10.
Int Urol Nephrol ; 50(8): 1535-1544, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29508174

RESUMO

PURPOSE: Podocytes, terminal differentiation cell in glomerulu, are crucial to kidney-related diseases such as membranous nephropathy (MN). MN is characterized by podocyte injury and glomerular basement membrane thickening. This paper focused to investigate the expression of chemokine (C-X-C motif) ligand 12 (CXCL12) in MN patients and its possible role in podocyte injury. METHODS: Through the enzyme-linked immunosorbent assay, CXCL12 level in the serum and urine of MN patients was examined. Further, several assays of cell viability, apoptosis, quantitative real-time PCR and western blot were applied to explore the effects of CXCL12 in the model of podocyte injury. RESULTS: We found a significant increase of CXCL12 in serum and urine of MN patients, which indicated that CXCL12 may be involved in the progression of MN. And in vitro C5b-9-induced podocyte injury model, the proliferation of podocytes was inhibited whereas CXCL12/CXCR4 and phosphorylated STAT3 (p-STAT3) were increased. Silencing of CXCL12 remarkably promoted cell proliferation, inhibited cell apoptosis and suppressed CXCL12/CXCR4, p-STAT3 and caspase 3. Consistently, STAT3 inhibitor and berberine (a CXCL12 antagonist) also reduced CXCL12 treatment-induced apoptosis. CONCLUSIONS: All data suggested that silencing of CXCL12 had a protective effect on podocyte injury, which may be through inhibiting CXCL12/STAT3 signaling pathway.


Assuntos
Quimiocina CXCL12/genética , Complexo de Ataque à Membrana do Sistema Complemento/efeitos adversos , Inativação Gênica , Glomerulonefrite Membranosa/genética , Podócitos/patologia , RNA/genética , Apoptose , Western Blotting , Sobrevivência Celular , Células Cultivadas , Quimiocina CXCL12/biossíntese , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Glomerulonefrite Membranosa/metabolismo , Glomerulonefrite Membranosa/patologia , Humanos , Fatores Imunológicos/efeitos adversos , Glomérulos Renais/efeitos dos fármacos , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Podócitos/efeitos dos fármacos , Podócitos/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais
11.
PLoS One ; 13(3): e0194298, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29529067

RESUMO

Rapid repair of vascular injury is an important prognostic factor for electrical burns. This repair is achieved mainly via stromal cell-derived factor (SDF)-1α promoting the mobilization, chemotaxis, homing, and targeted differentiation of bone marrow mesenchymal stem cells (BMSCs) into endothelial cells. Forming a concentration gradient from the site of local damage in the circulation is essential to the role of SDF-1α. In a previous study, we developed reactive oxygen species (ROS)-sensitive PPADT nanoparticles containing SDF-1α that could degrade in response to high concentration of ROS in tissue lesions, achieving the goal of targeted SDF-1α release. In the current study, a rat vascular injury model of electrical burns was used to evaluate the effects of targeted release of SDF-1α using PPADT nanoparticles on the chemotaxis of BMSCs and the repair of vascular injury. Continuous exposure to 220 V for 6 s could damage rat vascular endothelial cells, strip off the inner layer, significantly elevate the local level of ROS, and decrease the level of SDF-1α. After injection of Cy5-labeled SDF-1α-PPADT nanoparticles, the distribution of Cy5 fluorescence suggested that SDF-1α was distributed primarily at the injury site, and the local SDF-1α levels increased significantly. Seven days after injury with nanoparticles injection, aggregation of exogenous green fluorescent protein-labeled BMSCs at the injury site was observed. Ten days after injury, the endothelial cell arrangement was better organized and continuous, with relatively intact vascular morphology and more blood vessels. These results showed that SDF-1α-PPADT nanoparticles targeted the SDF-1α release at the site of injury, directing BMSC chemotaxis and homing, thereby promoting vascular repair in response to electrical burns.


Assuntos
Queimaduras por Corrente Elétrica/metabolismo , Queimaduras por Corrente Elétrica/patologia , Quimiocina CXCL12/biossíntese , Quimiotaxia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Nanopartículas , Espécies Reativas de Oxigênio/metabolismo , Animais , Biomarcadores , Biópsia , Queimaduras por Corrente Elétrica/tratamento farmacológico , Modelos Animais de Doenças , Masculino , Camundongos , Ratos , Cicatrização
12.
Immunity ; 48(2): 286-298.e6, 2018 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-29396162

RESUMO

Glucocorticoids are steroid hormones with strong anti-inflammatory and immunosuppressive effects that are produced in a diurnal fashion. Although glucocorticoids have the potential to induce interleukin-7 receptor (IL-7R) expression in T cells, whether they control T cell homeostasis and responses at physiological concentrations remains unclear. We found that glucocorticoid receptor signaling induces IL-7R expression in mouse T cells by binding to an enhancer of the IL-7Rα locus, with a peak at midnight and a trough at midday. This diurnal induction of IL-7R supported the survival of T cells and their redistribution between lymph nodes, spleen, and blood by controlling expression of the chemokine receptor CXCR4. In mice, T cell accumulation in the spleen at night enhanced immune responses against soluble antigens and systemic bacterial infection. Our results reveal the immunoenhancing role of glucocorticoids in adaptive immunity and provide insight into how immune function is regulated by the diurnal rhythm.


Assuntos
Ritmo Circadiano/fisiologia , Glucocorticoides/farmacologia , Receptores CXCR4/fisiologia , Receptores de Interleucina-7/fisiologia , Linfócitos T/imunologia , Animais , Células Cultivadas , Quimiocina CXCL12/biossíntese , Feminino , Memória Imunológica , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptores de Glucocorticoides/fisiologia
13.
Leukemia ; 32(6): 1445-1457, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29483712

RESUMO

CD69 regulates lymphocyte egress from the thymus and lymph nodes through cis-interactions and the downregulation of surface sphingosine-1-phosphate (S1P) receptor-1 (S1P1). However, its role in the regulation of cell egress from bone marrow has not been extensively studied. We show here that CD69 targeting induced rapid and massive mobilization of BM leukocytes, which was inhibited by desensitization to S1P with FTY720. This mobilization was reproduced with anti-human CD69 mAb treatment of mice expressing human CD69. In this strain, the mobilization occurred to the same extent as that induced by AMD3100. The anti-human CD69 treatment highly increased LSK and CLP cell proliferation and numbers, both in the periphery and in the BM, and also augmented S1P1 and CXCR4 expression. Additionally, increased mTOR, p70S6K, S6, and 4E-BP1 phosphorylation was detected after in vivo anti-CD69 treatment in the bone marrow. Importantly, mTOR inhibition with rapamycin inhibited anti-huCD69-induced mobilization of hematopoietic stem and progenitor cells (HSPCs). Together, our results indicated that CD69 targeting induces not only mobilization but also high proliferation of HSPCs, and thus is crucial for precursor cell replenishment over time. These results suggest that anti-CD69 mAbs are putative novel candidates for mobilization strategies.


Assuntos
Mobilização de Células-Tronco Hematopoéticas/métodos , Células-Tronco Hematopoéticas/fisiologia , Lectinas Tipo C/antagonistas & inibidores , Proteínas de Membrana/fisiologia , Monoéster Fosfórico Hidrolases/fisiologia , Serina-Treonina Quinases TOR/fisiologia , Animais , Anticorpos Monoclonais/farmacologia , Antígenos CD , Antígenos de Diferenciação de Linfócitos T , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Quimiocina CXCL12/biossíntese , Feminino , Cloridrato de Fingolimode/farmacologia , Compostos Heterocíclicos/farmacologia , Masculino , Camundongos
14.
Radiother Oncol ; 126(1): 125-131, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29061496

RESUMO

INTRODUCTION: Preclinical and clinical data suggest that the chemokine pathway governed by SDF-1 and CXCR4 contributes to a resistant phenotype. This retrospective biomarker study aims to explore the specific prognostic value of SDF-1 and CXCR4 expression in locally advanced head and neck squamous cell carcinomas (HNSCC) treated with primary radiochemotherapy (RT-CT). MATERIAL AND METHODS: Biopsies from 141 HNSCC tumours of the oral cavity, oropharynx and hypopharynx were evaluated for SDF-1 and CXCR4 expression by immunofluorescence. SDF-1 and CXCR4 expression was correlated with clinico-pathological characteristics and outcome after RT-CT. RESULTS: Patients with tumours exhibiting overexpression of intracellular SDF-1 and CXCR4 have a higher risk for loco-regional relapse and a worse overall survival after RT-CT (multivariate analysis, hazard ratio 2.33, CI [1.18-4.62], p = 0.02 and hazard ratio 2.02, CI [1.13-3.59], p = 0.02, respectively). Similar results were observed when only the subgroup of HPV DNA negative patients were analysed (hazard ratio 2.23 and 2.16, p = 0.02 and p = 0.01, respectively). CONCLUSIONS: Our data support the importance of SDF-1 and CXCR4 expression for loco-regional control and overall survival in HNSCC after primary radiochemotherapy. Prospective multivariate validation and further studies into CXCR4 inhibition to overcome radiation resistance are warranted.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/terapia , Quimiocina CXCL12/biossíntese , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/terapia , Receptores CXCR4/biossíntese , Idoso , Biomarcadores Tumorais/biossíntese , Carcinoma de Células Escamosas/tratamento farmacológico , Carcinoma de Células Escamosas/radioterapia , Quimiorradioterapia , Feminino , Neoplasias de Cabeça e Pescoço/tratamento farmacológico , Neoplasias de Cabeça e Pescoço/radioterapia , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/metabolismo , Recidiva Local de Neoplasia/patologia , Estadiamento de Neoplasias , Prognóstico , Estudos Prospectivos , Estudos Retrospectivos , Carcinoma de Células Escamosas de Cabeça e Pescoço , Taxa de Sobrevida
15.
J Am Heart Assoc ; 6(11)2017 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-29114002

RESUMO

BACKGROUND: Activated cardiac fibroblasts (CFs), preglomerular vascular smooth muscle cells (PGVSMCs), and glomerular mesangial cells (GMCs) proliferate, cause hypertrophy, and produce collagen; in this way, activated CFs contribute to cardiac fibrosis, and activated PGVSMCs and GMCs promote renal fibrosis. In heart and kidney diseases, SDF-1α (stromal cell-derived factor 1α; endogenous CXCR4 [C-X-C motif chemokine receptor 4] receptor agonist) levels are often elevated; therefore, it is important to know whether and how the SDF-1α/CXCR4 axis activates CFs, PGVSMCs, or GMCs. METHODS AND RESULTS: Here we investigated whether SDF-1α activates CFs, PGVSMCs, and GMCs to proliferate, hypertrophy, or produce collagen. DPP4 (dipeptidyl peptidase 4) inactivates SDF-1α and previous experiments show that growth-promoting peptides have greater effects in cells from genetically-hypertensive animals. Therefore, we performed experiments in the absence and presence of sitagliptin (DPP4 inhibitor) and in cells from normotensive Wistar-Kyoto rats and spontaneously hypertensive rats. Our studies show (1) that spontaneously hypertensive and Wistar-Kyoto rat CFs, PGVSMCs, and GMCs express CXCR4 receptors and DPP4 activity; (2) that chronic treatment with physiologically relevant concentrations of SDF-1α causes concentration-dependent increases in the proliferation (cell number) and hypertrophy (3H-leucine incorporation) of and collagen production (3H-proline incorporation) by CFs, PGVSMCs, and GMCs; (3) that sitagliptin augments these effects of SDF-1α; (4) that interactions between SDF-1α and sitagliptin are greater in spontaneously hypertensive rat cells; (5) that CXCR4 antagonism (AMD3100) blocks all effects of SDF-1α; and (6) that SDF-1α/CXCR4 signal transduction likely involves the RACK1 (receptor for activated C kinase 1)/Gßγ/PLC (phospholipase C)/PKC (protein kinase C) signaling complex. CONCLUSIONS: The SDF-1α/CXCR4 axis drives proliferation and hypertrophy of and collagen production by CFs, PGVSMCs, and GMCs, particularly in cells from genetically hypertensive animals and when DPP4 is inhibited.


Assuntos
Cardiomiopatia Hipertrófica/genética , Quimiocina CXCL12/genética , Colágeno/biossíntese , Regulação da Expressão Gênica , Células Mesangiais/patologia , Músculo Liso Vascular/patologia , Miocárdio/patologia , Animais , Western Blotting , Cardiomiopatia Hipertrófica/metabolismo , Cardiomiopatia Hipertrófica/patologia , Proliferação de Células , Células Cultivadas , Quimiocina CXCL12/biossíntese , Modelos Animais de Doenças , Fibroblastos/metabolismo , Fibroblastos/patologia , Glomérulos Renais/metabolismo , Glomérulos Renais/patologia , Masculino , Células Mesangiais/metabolismo , Microvasos/metabolismo , Microvasos/patologia , Músculo Liso Vascular/metabolismo , Miocárdio/metabolismo , RNA/genética , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Reação em Cadeia da Polimerase em Tempo Real , Receptores CXCR4/biossíntese , Receptores CXCR4/genética , Artéria Renal/metabolismo , Artéria Renal/patologia
16.
Proc Natl Acad Sci U S A ; 114(39): 10455-10460, 2017 09 26.
Artigo em Inglês | MEDLINE | ID: mdl-28900008

RESUMO

Antiangiogenic therapy with antibodies against VEGF (bevacizumab) or VEGFR2 (ramucirumab) has been proven efficacious in colorectal cancer (CRC) patients. However, the improvement in overall survival is modest and only in combination with chemotherapy. Thus, there is an urgent need to identify potential underlying mechanisms of resistance specific to antiangiogenic therapy and develop strategies to overcome them. Here we found that anti-VEGFR2 therapy up-regulates both C-X-C chemokine ligand 12 (CXCL12) and C-X-C chemokine receptor 4 (CXCR4) in orthotopic murine CRC models, including SL4 and CT26. Blockade of CXCR4 signaling significantly enhanced treatment efficacy of anti-VEGFR2 treatment in both CRC models. CXCR4 was predominantly expressed in immunosuppressive innate immune cells, which are recruited to CRCs upon anti-VEGFR2 treatment. Blockade of CXCR4 abrogated the recruitment of these innate immune cells. Importantly, these myeloid cells were mostly Ly6Clow monocytes and not Ly6Chigh monocytes. To selectively deplete individual innate immune cell populations, we targeted key pathways in Ly6Clow monocytes (Cx3cr1-/- mice), Ly6Chigh monocytes (CCR2-/- mice), and neutrophils (anti-Ly6G antibody) in combination with CXCR4 blockade in SL4 CRCs. Depletion of Ly6Clow monocytes or neutrophils improved anti-VEGFR2-induced SL4 tumor growth delay similar to the CXCR4 blockade. In CT26 CRCs, highly resistant to anti-VEGFR2 therapy, CXCR4 blockade enhanced anti-VEGFR2-induced tumor growth delay but specific depletion of Ly6G+ neutrophils did not. The discovery of CXCR4-dependent recruitment of Ly6Clow monocytes in tumors unveiled a heretofore unknown mechanism of resistance to anti-VEGF therapies. Our findings also provide a rapidly translatable strategy to enhance the outcome of anti-VEGF cancer therapies.


Assuntos
Inibidores da Angiogênese/farmacologia , Neoplasias Colorretais/terapia , Monócitos/imunologia , Neutrófilos/imunologia , Receptores CXCR4/metabolismo , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Animais , Anticorpos Monoclonais/farmacologia , Anticorpos Monoclonais Humanizados , Antígenos Ly/metabolismo , Bevacizumab/farmacologia , Proliferação de Células , Quimiocina CXCL12/biossíntese , Compostos Heterocíclicos/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores CXCR4/antagonistas & inibidores , Receptores CXCR4/biossíntese , Células Tumorais Cultivadas
17.
Wound Repair Regen ; 25(4): 652-664, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28783870

RESUMO

C-X-C chemokine receptor type 4 (CXCR4) is an alpha-chemokine receptor specific for stromal cell-derived factor 1 (SDF-1 also called CXCL12). The antagonist of CXCR4 can mobilize CD34+ cells and hematopoietic stem cells from bone marrow within several hours, and it has an efficacy on diabetes ulcer through acting on the SDF-1/CXCR4 axis. In this study, we investigated for the first time whether the antagonist of CXCR4 (Plerixafor/AMD3100) delivered on acellular dermal matrix (ADM) may accelerate diabetes-impaired wound healing. ADM scaffolds were fabricated from nondiabetic mouse skin through decellularization processing and incorporated with AMD3100 to construct ADM-AMD3100 scaffold. Full-thickness cutaneous wound in streptozotocin (STZ)-induced diabetic mice were treated with ADM, AMD3100, or ADM-AMD3100. 21 days after treatment, wound closure in ADM-AMD3100-treated mice was more complete than ADM group and AMD3100 group, and it was accompanied by thicker collagen formation. Correspondingly, diabetic mice treated with ADM-AMD3100 demonstrated prominent neovascularization (higher capillary density and vascular smooth muscle actin), which were accompanied by up-regulated mRNA levels of SDF-1 and enhanced migration of CXCR4 in the granulation tissue. Our results demonstrate that ADM scaffold provide perfect niche for loading AMD3100 and ADM-AMD3100 is a promising method for diabetic wound healing mainly by increasing expression of SDF-1 and enhancing migration of CXCR4-positive cells.


Assuntos
Derme Acelular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Quimiocina CXCL12/biossíntese , Receptores CXCR4/antagonistas & inibidores , Cicatrização/efeitos dos fármacos , Ferimentos e Lesões/tratamento farmacológico , Animais , Diabetes Mellitus Experimental , Tecido de Granulação , Compostos Heterocíclicos/farmacologia , Masculino , Camundongos , Receptores CXCR4/metabolismo , Relação Estrutura-Atividade , Ferimentos e Lesões/patologia
18.
J Immunol ; 199(7): 2343-2355, 2017 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-28842468

RESUMO

We investigated the involvement of CXCL12-CXCR4 interactions in human lymphohematopoiesis by coculture with telomerized human stromal cells. CXCR4 expression was low in CD34+CD38-CD45RA-CD10-CD7-CD19- immature hematopoietic stem/precursor cells (HSPCs) but higher in CD34+CD38-CD45RA+CD10+CD7+/-CD19- early lymphoid precursors and even higher in CD34+CD38+CD45RA+CD10+CD7-CD19+ pro-B cells. Inhibition of the effect of stromal cell-produced CXCL12 by an anti-CXCR4-blocking Ab suppressed the generation of CD45RA+CD10-CD7+CD19- early T lymphoid precursors (ETPs) and CD45RA+CD10+CD7-CD19+/- B lymphoid precursors on stromal cells, but it did not affect the generation of ETPs in conditioned medium of stromal cell cultures. Replating assays showed that contact with stromal cells was critical for HSPC-derived CD45RA+CD10+CD7-CD19- B lineage-biased precursors to differentiate into CD19+ pro-B cells, which was suppressed by the anti-CXCR4 Ab. Conversely, HSPC-derived ETPs possessed T and B lymphoid and monocytic differentiation potential; stromal cell contact was not required for their growth but rather promoted B lymphoid differentiation. The anti-CXCR4 Ab did not affect the growth of ETPs in conditioned medium, but it suppressed their B lymphoid differentiation on stromal cells. CD14-CD11c-HLA-DR+CD123highCD303+ plasmacytoid dendritic cells developed from HSPCs and ETPs exclusively in contact with stromal cells, which was suppressed by the anti-CXCR4 Ab. These data indicate that CXCL12 plays an essential role in stromal cell contact-mediated B lymphoid and plasmacytoid dendritic cell differentiation from immature hematopoietic and early T lymphoid precursors with a multilineage differentiation potential, but it does not participate in contact-independent generation of early T lymphoid precursors.


Assuntos
Linfócitos B/fisiologia , Diferenciação Celular , Quimiocina CXCL12/metabolismo , Células Dendríticas/fisiologia , Linfócitos/fisiologia , Receptores CXCR4/metabolismo , Linfócitos T/fisiologia , Antígenos CD19/genética , Antígenos CD34/genética , Células da Medula Óssea/citologia , Diferenciação Celular/imunologia , Linhagem da Célula , Quimiocina CXCL12/antagonistas & inibidores , Quimiocina CXCL12/biossíntese , Quimiocina CXCL12/imunologia , Técnicas de Cocultura , Meios de Cultivo Condicionados/farmacologia , Hematopoese , Humanos , Imunofenotipagem , Receptores CXCR4/genética , Receptores CXCR4/imunologia , Transdução de Sinais/imunologia , Células Estromais/efeitos dos fármacos , Células Estromais/fisiologia
19.
Eur Rev Med Pharmacol Sci ; 21(3 Suppl): 62-66, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28745789

RESUMO

OBJECTIVE: This study sought to identify the suitable cell culture conditions for the in vitro-induced differentiation of human embryonic stem cells (hESCs) into retinal vascular tissue cell types. MATERIALS AND METHODS: To do this, we established four treatment groups. Group A was designed to culture hESCs in a three-dimensional system. The feeder cells and leukemia inhibitory factor (LIF) were removed in Group B. In group C, hESCs were cultured with a variety of pro-angiogenic growth factors. In group D, hESCs were cultured with intact retinal support cells and extracellular matrix. On days 15 and 30, the expression of platelet endothelial cell adhesion molecule 1 (PECAM1), α-smooth muscle actin (αSMA), and macrophage marker F4/80 were detected by immunofluorescence staining. ELISA was used to detect the expression of stromal cell-derived factor-1 (SDF-1). RESULTS: At both 15 and 30 day timepoints, the highest PECAM1, αSMA, and F4/80 positive rates and SDF-1 expression levels were observed in group D, followed by group C, group B, with group A presenting the lowest expression of these proteins (p<0.05). Also, group D showed obvious angiogenesis structures. CONCLUSIONS: Our study indicates that hESCs can differentiate into retinal vascular-like structures. The presence of intact retinal support cells, a variety of cytokines, and extracellular matrix components were essential to facilitate this differentiation.


Assuntos
Técnicas de Cultura de Células , Células-Tronco Embrionárias Humanas/citologia , Vasos Retinianos/citologia , Actinas/genética , Diferenciação Celular , Células Cultivadas , Quimiocina CXCL12/biossíntese , Quimiocina CXCL12/genética , Humanos , Neovascularização Fisiológica/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética
20.
Tumour Biol ; 39(6): 1010428317706206, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28621237

RESUMO

C-X-C chemokine receptor type 4 and stromal cell-derived factor-1 were proven to play important roles in several types of cancer and in many biological processes connected with tumor growth, invasion, angiogenesis, and metastasis. However, the clinical significance of C-X-C chemokine receptor type 4 and stromal cell-derived factor-1 expression in colorectal cancer remains inaccurate. The purpose of this systematic meta-analysis is to investigate the role of C-X-C chemokine receptor type 4 and stromal cell-derived factor-1 as prognostic factors for survival and the association between C-X-C chemokine receptor type 4/ stromal cell-derived factor-1 and clinicopathology in colorectal cancer. Databases including PubMed, EMBASE, and Cochrane Library were searched for relevant literatures updated till January 2017. Review Manager 5.3 was used for data analysis. In our meta-analysis, C-X-C chemokine receptor type 4 expression is related to tumor-node-metastasis stage, tumor differentiation, liver metastasis, lymph node metastasis, distant metastasis, and diagnosis, and no correlation of C-X-C chemokine receptor type 4 expression with tumor size, gender, preoperative carcinoembryonic antigen, age, or vascular invasion has been observed. Stromal cell-derived factor-1 expression has no relationship with tumor-node-metastasis stage, lymph node metastasis, vascular invasion, age, gender, distant metastasis, or diagnosis. The expression of stromal cell-derived factor-1 has association with tumor differentiation. Moreover, the pooled hazard ratio for disease-free survival/overall survival showed that overexpression of C-X-C chemokine receptor type 4/stromal cell-derived factor-1 reduced disease-free survival/overall survival in colorectal cancer. Therefore, High expression of C-X-C chemokine receptor type 4/stromal cell-derived factor-1 which is essential in tumor progression can predict poor survival that may provide more advance prognostic clues to colorectal cancer patients.


Assuntos
Biomarcadores Tumorais/biossíntese , Quimiocina CXCL12/biossíntese , Neoplasias Colorretais/genética , Receptores CXCR4/biossíntese , Biomarcadores Tumorais/genética , Quimiocina CXCL12/genética , Neoplasias Colorretais/patologia , Intervalo Livre de Doença , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Metástase Linfática , Masculino , Estadiamento de Neoplasias , Prognóstico , Receptores CXCR4/genética
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