Nonspecific low back pain is closely associated with afferent nerve ingrowth into degenerated IVDs and increasing the inflammatory response. Members of the class 3 semaphorins signal their response through two prominent receptors; the NRP (Neuropilin-1) and the Plexin A. Sema3A (Semaphorin3A) is primarily known for their role in modulating neuronal survival as well as neurite outgrowth and guidance via regulation of Sema3A-NRP-1-plexinA signal pathway. Also, sema3A is shown to be conductive to innervate the inner painful degenerated IVDs (Intervertebral discs). Furthermore, sema3A is thought to act as a barrier to endothelial cells survival and migration on vascular endothelial growth factor (VEGF) and inhibition of KLF5-induced (Krüppel-like factor 5) inflammatory mediators within degenerated IVDs. Therefore, Sema3A produce a new perspective of dual-action therapeutic agent for attenuating the regulator of innervation and angiogenesis into degenerated IVDs and inhibition of KLF5-induced inflammation.
Endothelial Cells ; Humans ; Low Back Pain ; Neuropilin-1 ; Semaphorin-3A ; Vascular Endothelial Growth Factor A

Neuropilins (NRP1 and NRP2) are multifunctional receptor proteins that are involved in nerve, blood vessel, and tumor development. NRP1 was first found to be expressed in neurons, but subsequent studies have demonstrated its surface expression in cells from the endothelium and lymph nodes. NRP1 has been demonstrated to be involved in the occurrence and development of a variety of cancers. NRP1 interacts with various cytokines, such as vascular endothelial growth factor family and its receptor and transforming growth factor β1 and its receptor, to affect tumor angiogenesis, tumor proliferation, and migration. In addition, NRP1+ regulatory T cells (Tregs) play an inhibitory role in tumor immunity. High numbers of NRP1+ Tregs were associated with cancer prognosis. Targeting NRP1 has shown promise, and antagonists against NRP1 have had therapeutic efficacy in preliminary clinical studies. NRP1 treatment modalities using nanomaterials, targeted drugs, oncolytic viruses, and radio-chemotherapy have gradually been developed. Hence, we reviewed the use of NRP1 in the context of tumorigenesis, progression, and treatment.
Humans ; Neoplasms/drug therapy* ; Neovascularization, Pathologic ; Neuropilin-1 ; Vascular Endothelial Growth Factor A

OBJECTIVETo investigate the effect of RNA interference (RNAi) targeting NRP-1 gene on the growth and proliferation of nasopharyngeal carcinoma (NPC) CNE-2Z cells.

METHODSShort hairpin RNA (shRNA) plasmids targeting NRP-1 were designed and synthesized. These plasmids were respectively transfected into human NPC CNE-2Z cells and xenograft tumors in nude mice. The expressions of Fluorescein-labeled plasmids in NPC CNE-2Z cells and xenograft tumors were observed by fluorescence microscopy. Cell proliferation was detected by MTT assay. The inhibitory effects on target genes were evaluated with RT-PCR and Western blotting, respectively. The inhibitory effect of plasmids with the most effective sequence on xenograft tumors in nude mice was observed.

RESULTSCNE-2Z cell proliferation was significantly inhibited by NRP-1/shRNA silencing. RT-PCR showed NRP-1 mRNA expression was significantly decreased. Western blotting demonstrated the NRP-1/shRNA silencing can effectively inhibit the expression of target proteins in CNE-2Z cells. After six weeks, there were significant differences in the mean tumor volumes in nude mice between plasmid group and negative control group [(0.599±0.002) vs (1.141±0.013) cm(3), P<0.05] or blank control group [(0.599±0.002) vs (1.165±0.308) cm(3), P<0.05], and the inhibitory rate of tumor growth was 48.6%.

CONCLUSIONRNA interference targeting NRP-1 can remarkably inhibit the growth of CNE-2Z cells in vitro and in vivo.

Animals ; Apoptosis ; Carcinoma ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Gene Targeting ; Humans ; Mice ; Mice, Nude ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neuropilin-1 ; metabolism ; Plasmids ; RNA Interference ; RNA, Small Interfering ; Transfection

This study was aimed to investigate the expression of neuropilin-1 (NRP-1) and NRP-2 mRNA in myeloid leukemia cells and the effect of NRP-1 on cell proliferation and migration. The expressions of NRP-1 and NRP-2 mRNA in bone marrow mononuclear cells of 24 patients with acute myeloid leukemia and in 7 myeloid leukemic cell lines (HL-60, KGIa, NB4, U937, HEL MEG01 and K562) were detected by RT-PCR. The effects of NRP-1 interfered with siRNA on proliferation and migration in leukemic cell line HEL were examined by MTT and migration test. The results showed that the expression of NRP-1 mRNA was found in bone marrow mononuclear cells (BMMNCS) of 24 AML patients, the positive rate was 100% and significantly higher than that in control group (positive rate 67%). The expressions of NRP-2 mRNA were seen in 79% AML patients and in 67% health control, there was no significant difference between them. The increased NRP-1 expression was directly correlated with the blast percentage in both peripheral blood and bone marrow of AML patients (r=05, r=0.4, p<0.05). The expressions of NRP-1 and NRP-2 mRNA were observed in 6/7 and 3/7 myeloid leukemic cell lines respectively. After HEL cells were transfected with siRNA for 24 hours, the expression levels of NRP-1 mRNA and protein decreased obviously. Under VEGF action, the cell number in control group significantly increased, while the cell proliferation in interfered group had been not changed. After being transfected for 24 hours, the migration in interfered group decreased significantly. It is concluded that the higher level of NRP-1 mRNA is expressed in bone marrow mononuclear cells of leukemia patients and plays a pivotal role in proliferation and migration of myeloid leukemic cells. Inhibition of NRP-1 functions may provide a new therapeutic strategy for AML.
Adolescent ; Adult ; Aged ; Cell Movement ; Cell Proliferation ; Child ; Female ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; Neuropilin-1 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; pharmacology ; Tumor Cells, Cultured ; Young Adult

Psoriasis is a chronic, recurrent, heterogeneous, cutaneous inflammatory skin disease for which there is no cure. It affects approximately 7.5 million people in the United States. Currently, several biologic agents that target different molecules implicated in the pathogenic processes of psoriasis are being assessed in diverse clinical studies. However, relapse usually occurs within weeks or months, meaning there is currently no cure for psoriasis. Therefore, recent studies have discovered diverse new potential treatments for psoriasis: inhibitors of bacteria such as Staphylococcus aureus, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and neuropilin 1 (NRP1). A promising approach that has recently been described involves modifying antimicrobial peptides to develop new cutaneous anti-bacterial agents that target inflammatory skin disease induced by Staphylococcus. Increased expression of TRAIL and its death receptors DR4 and DR5 has been implicated in the pathogenesis of plaque psoriasis. In addition, TRAIL has the ability to inhibit angiogenesis by inducing endothelial cell death and by negative regulation of VEGF-induced angiogenesis via caspase-8-mediated enzymatic and non-enzymatic functions. Since NRP1 regulates angiogenesis induced by multiple signals, including VEGF, ECM and semaphorins, and also initiates proliferation of keratinocytes through NF-κB signaling pathway in involved psoriatic skin, targeting NRP1 pathways may offer numerous windows for intervention in psoriasis. In this review, we will focus on the current knowledge about the emerging role of synthetic antimicrobial peptides, TRAIL and NRP1 blocking peptides in the pathogenesis and treatment of psoriasis.
Anti-Bacterial Agents ; Bacteria ; Biological Factors ; Endothelial Cells ; Keratinocytes ; Necrosis ; Neuropilin-1 ; Peptides ; Psoriasis ; Receptors, Death Domain ; Recurrence ; Semaphorins ; Skin ; Skin Diseases ; Staphylococcus ; Staphylococcus aureus ; Therapeutic Uses ; TNF-Related Apoptosis-Inducing Ligand ; United States ; Vascular Endothelial Growth Factor A

PURPOSE : The purpose of this study was to examine the expression of various angiogenic factors during osteoblastic differentiation of periostealderived cells and the effects of osteogenic inductive medium of periosteal-derived cells on the proliferation of endothelial progenitor cells. MATERIALS AND METHODS : Periosteal-derived cells were obtained from mandibular periosteums and introduced into the cell culture. After passage 3, the cells were divided into two groups and cultured for 21 days. In one group, the cells were cultured in the DMEM supplemented with osteogenic inductive agent, including 50g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate. In the other group, they were cultured in DMEM supplemented without osteogenic inductive agent. VEGF isoforms, VEGFR-1, VEGFR-2, and neuropilin-1 mRNA expression was observed. Human umbilical cord blood-derived endothelial progenitor cell proliferation was also observed. RESULTS : The expression of VEGF isoforms was higher in osteogenic inductive medium than in non-osteogenic inductive medium. The expression of VEGFR-2 was also higher in osteogenic inductive medium than in non-osteogenic inductive medium. However, the expression of VEGFR-1 and neuropilin-1 was similar in both osteogenic inductive medium and non-osteogenic inductive medium. In addition, conditioned medium from differentiated periosteal-derived cells stimulated human umbilical cord blood-derived endothelial progenitor cell numbers compared to conditioned medium from non-differentiated periosteal-derived cells. CONCLUSION : These results suggest that in vitro osteoblastic differentiation of periosteal-derived cells has angiogenic capacity to support endothelial progenitor cell numbers.
Angiogenesis Inducing Agents ; Ascorbic Acid ; Cell Culture Techniques ; Culture Media, Conditioned ; Dexamethasone ; Durapatite ; Humans ; Neuropilin-1 ; Osteoblasts ; Periosteum ; Protein Isoforms ; RNA, Messenger ; Stem Cells ; Umbilical Cord ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-2

Neuropilin 1 (NP1) is a part of essential receptor complexes mediating both semaphorin3A (SEMA3A) and vascular endothelial growth factor (VEGF) which is one of important mediators involved in the pathogenesis of asthma. Therefore, it is possible that SEMA3A plays a role in the pathogenesis of asthma through attenuation of VEGF-mediated effects. In the present study, we aimed to evaluate expression levels of SEMA3A and NP1 using induced sputum of asthmatics and a murine model of asthma. Firstly, SEMA3A and NP1 expressions in induced sputum of asthmatics and SEMA3A and NP1 expression on bronchoalveolar lavage (BAL) cells and lung homogenates of asthmatic mice were determined. Then we evaluated the immunolocalization of VEGF receptor 1 (VEGFR1), VEGF receptor 2 (VEGFR2), and NP1 expressions on asthmatic mice lung tissue and their subcellular distributions using fibroblast and BEAS2B cell lines. Sputum SEMA3A and NP1 expressions were significantly higher in asthmatics than controls. Similarly, SEMA3A and NP1 expressions on BAL cells and lung homogenates were significantly elevated in asthmatic mice compared to control mice. Immunohistochemical analysis showed that VEGFR1, VEGFR2, and NP1 expressions were also uniformly increased in asthmatic mice. Our observations suggest that SEMA3A and NP1 may play important roles in the pathogenesis of asthma.
Animals ; Asthma/metabolism/pathology/*physiopathology ; Bronchoalveolar Lavage Fluid/cytology ; Cell Line ; Disease Models, Animal ; Female ; Fibroblasts/metabolism ; *Gene Expression Regulation ; Immunohistochemistry ; Lung/metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Neuropilin-1/*genetics/metabolism ; Semaphorin-3A/*genetics/metabolism ; Sputum/metabolism ; Vascular Endothelial Growth Factor Receptor-1/metabolism ; Vascular Endothelial Growth Factor Receptor-2/metabolism

1, VEGFR-2, and neuropilin-1. ALL VEGF isoforms (VEGF(121), VEGF(165), VEGF(189), and VEGF(206)) expression was observed by RT-PCR analysis. VEGFR-1, VEGFR-2 and neuropilin-1 expression increased up to day 14, particularly during the early stage of mineralization. Our results suggest the involvement of direct VEGFs/VEGFRs system on periosteal-derived cells during early mineralization phase under high-dose of dexamethasone. These also suggest that VEGF might act as an autocrine growth molecule during osteoblastic differentiation of cultured human periosteal-derived cells.]]>
Ascorbic Acid ; Bone Development ; Bone Marrow ; Dexamethasone ; Durapatite ; Fractures, Bone ; Gene Expression ; Glycerophosphates ; Humans ; Neuropilin-1 ; Osteoblasts ; Osteocalcin ; Osteogenesis ; Phenotype ; Protein Isoforms ; Receptors, Vascular Endothelial Growth Factor ; Stromal Cells ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-1 ; Vascular Endothelial Growth Factor Receptor-2

Objective: To investigate the expression and function of plexin A1, a transmembrane protein involving cell survival and cell-to cell interaction, in the rheumatoid synoviocytes. Methods: Immunohistochemical staining using anti-plexin A1 antibody was performed in the synovium of rheumatoid arthritis (RA) patients. The plexin A1 expression in cultured fibroblast-like synovioytes (FLS) was also examined by Western blot analysis and immunocytochemistry. Cell viability was determined by CCK-8 assay. Deficiency of plexin A1 was established by the method of short interfering RNA (siRNA). The productions of interleukin-6 (IL-6) and monocytes chemotactic protein-1 (MCP-1) were measured in culture supernatants by ELISA. Results: Plexin A1 was highly expressed in the lining layer of synovium and cultured FLS of RA patients. In RA FLS, basal expression of plexin A1 was higher than osteoarthritis FLS. On immunocytochemical staining, plexin A1 was co-expressed with neuropilin-1 in RA FLS. Semaphorin 3A (10 to 200 ng/mL), a specific ligand for neuropilin-1/plexin A1 complex, did not affect viability of RA FLS. The down regulation of plexin A1 mRNA by siRNA did not cause cell death, either. Co-culture of FLS with RA T cells, isolated from peripheral blood or synovial fluid, caused an increase in the productions of IL-6 and MCP-1 from FLS, but which were blocked by down-regulating plexin A1 transcripts using siRNA method. Conclusion: These data suggest that enhanced expression of plexin A1 in RA FLS may elicit over-production of IL-6 and MCP-1, and thereby contribute to perpetuation of chronic inflammation in RA.
Arthritis, Rheumatoid ; Blotting, Western ; Cell Communication ; Cell Death ; Cell Survival ; Coculture Techniques ; Down-Regulation ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Inflammation ; Interleukin-6 ; Monocytes ; Neuropilin-1 ; Osteoarthritis ; RNA, Messenger ; RNA, Small Interfering ; Semaphorin-3A ; Sincalide ; Synovial Fluid ; Synovial Membrane ; T-Lymphocytes

OBJECTIVEThe purpose of the present study was to observe the changes in CD4+CD25+Nrp1+Treg cells after irradiation with different doses and explore the possible molecular mechanisms involved.

METHODSICR mice and mouse lymphoma cell line (EL-4 cells) was used. The expressions of CD4, CD25, Nrp1, calcineurin and PKC-α were detected by flow cytometry. The expressions of TGF-β1, IL-10, PKA and cAMP were estimated with ELISA.

RESULTSAt 12 h after irradiation, the expression of Nrp1 increased significantly in 4.0 Gy group, compared with sham-irradiation group (P<0.05) in the spleen and thymus, respectively, when ICR mice received whole-body irradiation (WBI). Meanwhile the synthesis of Interleukin 10 (IL-10) and transforming growth factor-β1 (TGF-β1) increased significantly after high dose irradiation (HDR) (> or = 1.0 Gy). In addition, the expression of cAMP and PKA protein increased, while PKC-α, calcineurin decreased at 12h in thymus cells after 4.0 Gy X-irradiation. While TGF-β1 was clearly inhibited when the PLC-PIP2 signal pathway was stimulated or the cAMP-PKA signal pathway was blocked after 4.0 Gy X-irradiation, this did not limit the up-regulation of CD4+CD25+Nrp1+Treg cells after ionizing radiation.

CONCLUSIONThese results indicated that HDR might induce CD4+CD25+Nrp1+Treg cells production and stimulate TGF-β1 secretion by regulating signal molecules in mice.

Animals ; Calcineurin ; genetics ; metabolism ; Cyclic AMP ; metabolism ; Dose-Response Relationship, Radiation ; Female ; Gene Expression Regulation ; radiation effects ; Immunosuppression ; Interleukin-10 ; genetics ; metabolism ; Lymphocyte Subsets ; physiology ; Male ; Mice ; Neuropilin-1 ; genetics ; metabolism ; Phosphoinositide Phospholipase C ; genetics ; metabolism ; Protein Kinases ; genetics ; metabolism ; Signal Transduction ; Transforming Growth Factor beta ; genetics ; metabolism ; Whole-Body Irradiation ; adverse effects