Tertiary RNA Folding-Targeted Drug Screening Strategy Using a Protein Nanopore.

Bacterial riboswitch RNAs are attractive targets for novel antibiotics against antibiotic-resistant superbacteria. Their binding to cognate metabolites is essential for the regulation of bacterial gene expression. Despite the importance of RNAs as therapeutic targets, the development of RNA-targeted, small-molecule drugs is limited by current biophysical methods. Here, we monitored the specific interaction between the adenine-sensing riboswitch aptamer domain (ARS) and adenine at the single-molecule level using α-hemolysin (αHL) nanopores. During adenine-induced tertiary folding, adenine-bound ARS intermediates exhibited characteristic nanopore events, including a two-level ionic current blockade and a ∼ 5.6-fold longer dwell time than that of free RNA. In a proof-of-concept experiment, tertiary RNA folding-targeted drug screening was performed using a protein nanopore, which resulted in the discovery of three new ARS-targeting hit compounds from a natural compound library. Taken together, these results reveal that αHL nanopores are a valuable platform for ultrasensitive, label-free, and single-molecule-based drug screening against therapeutic RNA targets.

A preclinical trial of perventricular pulmonary valve implantation: Pericardial versus aortic porcine valves mounted on self-expandable stent.

Perventricular pulmonary valve implantation (PPVI) of a xenograft valve can be a less invasive technique that avoids cardiopulmonary bypass in patients who require pulmonary valve replacement. We compared the hemodynamics, durability, and histologic changes between two different xenogenic valves (pericardial vs. aortic valve porcine xenografts) implanted into the pulmonary valve position using a PPVI technique and evaluated the safety and efficacy of PPVI as a preclinical study. In 18 sheep, pericardial (group porcine pericardial [PP], n = 9) or aortic valve (group porcine aortic valve [PAV], n = 9) xenogenic porcine valves manufactured as a stented valve were implanted using a PPVI technique. The porcine tissues were decellularized, alpha-galactosidase treated, fixed with glutaraldehyde after space-filler treatment, and detoxified to improve durability. Hemodynamic and immunohistochemical studies were performed after the implantation; radiologic and histologic studies were performed after a terminal procedure. All stented valves were positioned properly after the implantation, and echocardiography and cardiac catheterization demonstrated good hemodynamic state and function of the valves. All the anti-α-Gal IgM and IgG titers were below 0.3 optical density. Computed tomography of extracted valves demonstrated no significant differences in the degree of calcification between the two groups (P = .927). Microscopic findings revealed a minimal amount of calcification and no significant infiltration of macrophage or T-cell in both groups, regardless of the implantation duration. The PPVI is a feasible technique. Both stented valves made of PP and PAV showed no significant differences in hemodynamic profile, midterm durability, and degree of degenerative dystrophic calcification.

Surface modification of solid-state nanopore by plasma-polymerized chemical vapor deposition of poly(ethylene glycol) for stable device operation.

Biopolymer adsorption onto a membrane is a significant issue in the reliability of solid-state nanopore devices, since it degrades the device performance or promotes device failure. In this work, a poly(ethylene glycol) (PEG) layer was coated on a silicon nitride (SiNx) membrane by plasma-polymerized vapor deposition to inhibit biopolymer adsorption. From optical observations, the deposited PEG layer demonstrated increased hydrophilicity and anti-adsorption property compared to the SiNx surface. Electrical properties of the PEG/SiNx nanopore were characterized, showing Ohmic behavior and a 6.3 times higher flicker noise power due to the flexible conformation of PEG in water. Antifouling performance of each surface was analyzed by measuring the average time from voltage bias to the first adsorption during DNA translocation experiments, where the modified surface enabled two times prolonged device operation. The time to adsorption was dependent on the applied voltage, implying adsorption probability was dominated by the electrophoretic DNA approach to the nanopore. DNA translocation behaviors on each surface were identified from translocation signals, as the PEG layer promoted unfolded and fast movement of DNA through the nanopore. This work successfully analyzed the effect of the PEG layer on DNA adsorption and translocation in solid-state nanopore experiments.

Electrical properties of graphene/In2O3 bilayer with remarkable uniformity as transparent conducting electrode.

A graphene/In2O3 bilayer (termed as GI-bilayer) is proposed as a transparent conducting electrode with remarkably improved areal-uniformity. To fabricate this new structure, an In2O3 layer with a thickness of less than 50 nm was grown by atomic layer deposition and then a graphene layer was grown by chemical vapor deposition and subsequently transferred onto the as-grown In2O3 layer. Electrical and optical properties of the GI-bilayer were systematically studied to verify effects of the underlying In2O3 layer. Hall measurements and following analysis showed a conductance enhancement of the GI-bilayer owing to p-type doping of graphene. Specifically, Raman analysis and ultraviolet photoelectron spectroscopy were performed to prove p-type doping of the graphene in the GI-bilayer. In addition, the GI-bilayer exhibited the significantly improved uniformity of the sheet resistance compared to that of a conventional monolayer of graphene. There was a duality on the role of the In2O3 underlayer in the GI-bilayer. It acted as a dopant layer to the graphene and lowered the sheet resistance from 863 to 510 Ω/sq as well as compensated microscale defects on graphene. More importantly, the In2O3 underlayer resulted in the extremely reduced standard deviation of sheet resistance from 150 to 7.5 Ω/sq over the area of 49 cm2.

Microfluidic electrochemical cell for in situ structural characterization of amorphous thin-film catalysts using high-energy X-ray scattering.

Porous, high-surface-area electrode architectures are described that allow structural characterization of interfacial amorphous thin films with high spatial resolution under device-relevant functional electrochemical conditions using high-energy X-ray (>50 keV) scattering and pair distribution function (PDF) analysis. Porous electrodes were fabricated from glass-capillary array membranes coated with conformal transparent conductive oxide layers, consisting of either a 40 nm-50 nm crystalline indium tin oxide or a 100 nm-150 nm-thick amorphous indium zinc oxide deposited by atomic layer deposition. These porous electrodes solve the problem of insufficient interaction volumes for catalyst thin films in two-dimensional working electrode designs and provide sufficiently low scattering backgrounds to enable high-resolution signal collection from interfacial thin-film catalysts. For example, PDF measurements were readily obtained with 0.2 Šspatial resolution for amorphous cobalt oxide films with thicknesses down to 60 nm when deposited on a porous electrode with 40 µm-diameter pores. This level of resolution resolves the cobaltate domain size and structure, the presence of defect sites assigned to the domain edges, and the changes in fine structure upon redox state change that are relevant to quantitative structure-function modeling. The results suggest the opportunity to leverage the porous, electrode architectures for PDF analysis of nanometre-scale surface-supported molecular catalysts. In addition, a compact 3D-printed electrochemical cell in a three-electrode configuration is described which is designed to allow for simultaneous X-ray transmission and electrolyte flow through the porous working electrode.

Detection of metal corrosion characteristics in chlorine solution using solid state nanopore.

Nanopore structures were originally proposed for detection of biomolecule translocation through nanometer-scale pores that perforate membranes by transient changes in ionic current. In this study, these changes are utilized to detect corrosion of different metals in aqueous chlorine media. The corrosion behaviors of Cu, Al, Ti, and Cr were analyzed by monitoring the changes in ion current resulting from ion concentration variations in solutions due to corrosion of the metals. We observed that the Cu layer passivated by CuO x was severely corroded when a drastic change of ion current was induced, from 10 to 30 nS to the level of 104 nS, as soon as the bias voltage was applied. In the case of Al passivated by thin AlO x , the conductance increased from 10-30 to 166 ± 52 nS and became saturated. Highly localized pitting corrosion was observed on the periphery of the nanopore, where the electrical field was most concentrated. Finally, we observed that Ti and Cr passivated by oxide showed long-term stability without corrosion in 1 M KCl in the pH range of 4-11.

DNA translocation through a nanopore in an ultrathin self-assembled peptide membrane.

Here, we explore the possibility of using peptide-based materials as a membrane in solid-state nanopore devices in an effort to develop a sequence-specific, programmable biological membrane platform. We use a recently developed tyrosine-mediated self-assembly peptide sheet. At the air/water interface, the 5mer peptide YFCFY self-assembles into a uniform and robust two-dimensional (2D) structure, and the peptide sheet is easily transferred to a low-noise glass substrate. The thickness of the peptide membrane can be adjusted to approximately 5 nm (or even to 2 nm) by an etching process, and the diameters of the peptide nanopores can be precisely controlled using a focused electron beam with an attuned spot size. The ionic current noise of the peptide nanopore is comparable to those of typical silicon nitride nanopores or multilayer 2D materials. Using this membrane, we successfully observe translocation of 1000 bp double-stranded DNA with a sufficient signal-to-noise ratio of ∼30 and an elongated translocation speed of ∼1 bp µs-1. Our results suggest that the self-assembled peptide film can be used as a sensitive nanopore membrane and employed as a platform for applying biological functionalities to solid-state substrates.

Incisional Ptosis Correction with Hidden Double Fold in Asian Patients.

BACKGROUND: Korean males and a few females desire to have larger eyes; however, they often wish to enlarge their eyes and conceal their double eyelids. This paper attempts to describe how to make the eyes bigger and brighter without showing double-fold eyelids. METHODS: The authors performed cosmetic ptosis correction in 121 cases from April 2013 to December 2017. All patients enrolled in this retrospective study underwent surgical procedures at the author's institutions. Patients were included that had mild-to-moderate degrees of ptosis and levator function greater than 5 mm, ages greater than 16 years, and no prior ptosis surgery. RESULTS: A successful outcome was achieved with this surgical approach in 113 (93.4%) patients. Complications potentially associated with ptosis surgery were not observed. DISCUSSION: A refined method of preoperative evaluation for incisional ptosis correction to conceal a double fold with no visible signs of surgery is described. Ptosis correction without the formation of double eyelids will result in skin hooding and visible scarring, and thus, it is recommended to lower the height of the double eyelids. The lower height of double eyelids can cover the incisional scar and make it appear there are no double eyelids. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Reduction of DNA Folding by Ionic Liquids and Its Effects on the Analysis of DNA-Protein Interaction Using Solid-State Nanopore.

DNA folding is not desirable for solid-state nanopore techniques when analyzing the interaction of a biomolecule with its specific binding sites on DNA since the signal derived from the binding site could be buried by a large signal from the folding of DNA nearby. To resolve the problems associated with DNA folding, ionic liquids (ILs), which are known to interact with DNA through charge-charge and hydrophobic interactions are employed. 1-n-butyl-3-methylimidazolium chloride (C4 mim) is found to be the most effective in lowering the incident of DNA folding during its translocation through solid-state nanopores (4-5 nm diameter). The rate of folding signals from the translocation of DNA-C4 mim is decreased by half in comparison to that from the control bare DNA. The conformational changes of DNA upon complexation with C4 mim are further examined using atomic force microscopy, showing that the entanglement of DNA which is common in bare DNA is not observed when treated with C4 mim. The stretching effect of C4 mim on DNA strands improves the detection accuracy of nanopore for identifying the location of zinc finger protein bound to its specific binding site in DNA by lowering the incident of DNA folding.

Sweet's syndrome: a clinical entity need to discriminate against acute haematogenous periprosthetic joint infection.

Sweet's syndrome (SS) or acute febrile neutrophilic dermatosis is an uncommon condition. It is characterized by fever, polymorphonuclear leukocytosis, painful erythematous cutaneous plaques, and dense dermal infiltrate of neutrophils without vasculitis at the site of skin lesions. Lesions in SS might enlarge and coalesce with increasing dermal oedema, resulting in pseudo-vesicular appearance mimicking joint infections. Here, a rare case of SS mimicking acute haematogenous periprosthetic infection in a 74-year-old woman with a history of total knee arthroplasty is reported. This report aims to elaborate clinical various manifestations of SS in a patient with a history of total knee arthroplasty. In addition, this report describes how to discriminate inflammation between SS and periprosthetic joint infection. Level of evidence V.

A novel role for flotillin-1 in H-Ras-regulated breast cancer aggressiveness.

Elevated expression and aberrant activation of Ras have been implicated in breast cancer aggressiveness. H-Ras, but not N-Ras, induces breast cell invasion. A crucial link between lipid rafts and H-Ras function has been suggested. This study sought to identify the lipid raft protein(s) responsible for H-Ras-induced tumorigenicity and invasiveness of breast cancer. We conducted a comparative proteomic analysis of lipid raft proteins from invasive MCF10A human breast epithelial cells engineered to express active H-Ras and non-invasive cells expressing active N-Ras. Here, we identified a lipid raft protein flotillin-1 as an important regulator of H-Ras activation and breast cell invasion. Flotillin-1 was required for epidermal growth factor-induced activation of H-Ras, but not that of N-Ras, in MDA-MB-231 triple-negative breast cancer (TNBC) cells. Flotillin-1 knockdown inhibited the invasiveness of MDA-MB-231 and Hs578T TNBC cells in vitro and in vivo. In xenograft mouse tumor models of these TNBC cell lines, we showed that flotillin-1 played a critical role in tumor growth. Using human breast cancer samples, we provided clinical evidence for the metastatic potential of flotillin-1. Membrane staining of flotillin-1 was positively correlated with metastatic spread (p = 0.013) and inversely correlated with patient disease-free survival rates (p = 0.005). Expression of flotillin-1 was associated with H-Ras in breast cancer, especially in TNBC (p < 0.001). Our findings provide insight into the molecular basis of Ras isoform-specific interplay with flotillin-1, leading to tumorigenicity and aggressiveness of breast cancer.

Probing the Small-Molecule Inhibition of an Anticancer Therapeutic Protein-Protein Interaction Using a Solid-State Nanopore.

Nanopore sensing is an emerging technology for the single-molecule-based detection of various biomolecules. In this study, we probed the anticancer therapeutic p53 transactivation domain (p53TAD)/MDM2 interaction and its inhibition with a small-molecule MDM2 antagonist, Nutlin-3, using low-noise solid-state nanopores. Although the translocation of positively charged MDM2 through a nanopore was detected at the applied negative voltage, this MDM2 translocation was almost completely blocked upon formation of the MDM2/GST-p53TAD complex owing to charge conversion. In combination with NMR data, the nanopore measurements showed that the addition of Nutlin-3 rescued MDM2 translocation, indicating that Nutlin-3 disrupted the MDM2/GST-p53TAD complex, thereby releasing MDM2. Taken together, our results reveal that solid-state nanopores can be a valuable platform for the ultrasensitive, picomole-scale screening of small-molecule drugs against protein-protein interaction (PPI) targets.

Visceral obesity in predicting oncologic outcomes of localized renal cell carcinoma.

PURPOSE: We investigate the clinicopathological features and prognostic significance of visceral obesity in patients with localized renal cell carcinoma. MATERIALS AND METHODS: This study included 706 patients with localized renal cell carcinoma who had undergone curative surgery between January 2003 and April 2012. Visceral, subcutaneous and total adipose tissue were measured based on preoperative computerized tomography of the umbilical region. Visceral adipose tissue percent was calculated using the formula, VAT% = [visceral adipose tissue/total adipose tissue] × 100. The association between clinicopathological factors and visceral obesity was examined. RESULTS: A higher VAT% at diagnosis was associated with older age at diagnosis, higher prevalence of diabetes and higher prevalence of former or current smoking status. The distribution of histological subtypes differed significantly among VAT% quartiles. The proportion of high grade tumors increased as VAT% increased (OR 1.023, 95% CI 1.000-1.126, p = 0.037). A U-shaped association between VAT% quartiles and the risk of disease recurrence was observed for all patients. Disease recurrence was significantly increased in the lowest (HR 3.198, 95% CI 1.765-10.040, p = 0.036) and highest (HR 4.760, 95% CI 2.937-13.210, p = 0.010) VAT% quartiles. CONCLUSIONS: Relative visceral obesity as assessed by VAT% was associated with clinicopathological characteristics of localized renal cell carcinoma. A U-shaped association between VAT% quartiles and risk of disease recurrence was observed among patients with localized renal cell carcinoma.

Synthesis of triphenylsulfonium triflate bound copolymer for electron beam lithography.

Photoacid generator (PAG) has been widely used as a key component in photoresist for high-resolution patterning with high sensitivity. A novel acrylic monomer, triphenylsulfonium salt methyl methacrylate (TPSMA), was synthesized and includes triphenylsulfonium triflate as a PAG. The poly(MMA-co-TPSMA) (PMT) as a polymer-bound PAG was synthesized with methyl methacrylate (MMA) and TPSMA for electron beam lithography. Characterization of PMT was carried out by NMR and FTIR. The molecular weight was analyzed by GPC. Thermal properties were studied using TGA and DSC. Thecharacterization results were in good agreement with corresponding chemical compositions and thermal stability. PMT was subsequently employed in electron beam lithography and its lithographic performance was confirmed by FE-SEM. This PMT was accomplished to improve the lithographic performance including sensitivity, line width roughness (LWR) and resolution. We found that PMT was capable of 20 nm negative tone patterns with better sensitivity than hydrogensilsesquioxane (HSQ) which is a conventional negative tone resist.

Nitric oxide releasing nanofiber stimulates revascularization in response to ischemia via cGMP-dependent protein kinase.

Nitric oxide (NO) promotes angiogenesis via various mechanisms; however, the effective transmission of NO in ischemic diseases is unclear. Herein, we tested whether NO-releasing nanofibers modulate therapeutic angiogenesis in an animal hindlimb ischemia model. Male wild-type C57BL/6 mice with surgically-induced hindlimb ischemia were treated with NO-releasing 3-methylaminopropyltrimethoxysilane (MAP3)-derived or control (i.e., non-NO-releasing) nanofibers, by applying them to the wound for 20 min, three times every two days. The amount of NO from the nanofiber into tissues was assessed by NO fluorometric assay. The activity of cGMP-dependent protein kinase (PKG) was determined by western blot analysis. Perfusion ratios were measured 2, 4, and 14 days after inducing ischemia using laser doppler imaging. On day 4, Immunohistochemistry (IHC) with F4/80 and gelatin zymography were performed. IHC with CD31 was performed on day 14. To determine the angiogenic potential of NO-releasing nanofibers, aorta-ring explants were treated with MAP3 or control fiber for 20 min, and the sprout lengths were examined after 6 days. As per either LDPI (Laser doppler perfusion image) ratio or CD31 capillary density measurement, angiogenesis in the ischemic hindlimb was improved in the MAP3 nanofiber group; further, the total nitrate/nitrite concentration in the adduct muscle increased. The number of macrophage infiltrations and matrix metalloproteinase-9 (MMP-9) activity decreased. Vasodilator-stimulated phosphoprotein (VASP), one of the major substrates for PKG, increased phosphorylation in the MAP3 group. MAP3 nanofiber or NO donor SNAP (s-nitroso-n-acetyl penicillamine)-treated aortic explants showed enhanced sprouting in an ex vivo aortic ring assay, which was partially abrogated by KT5823, a potent inhibitor of PKG. These findings suggest that the novel NO-releasing nanofiber, MAP3 activates PKG and promotes therapeutic angiogenesis in response to hindlimb ischemia.

Noise and its reduction in graphene based nanopore devices.

Ionic current fluctuations in graphene nanopore devices are a ubiquitous phenomenon and are responsible for degraded spatial and temporal resolution. Here, we descriptively investigate the impact of different substrate materials (Si and quartz) and membrane thicknesses on noise characteristics of graphene nanopore devices. To mitigate the membrane fluctuations and pin-hole defects, a SiNx membrane is transferred onto the substrate and a pore of approximately 70 nm in diameter is perforated prior to the graphene transfer. Comprehensive noise study reveals that the few layer graphene transferred onto the quartz substrate possesses low noise level and higher signal to noise ratio as compared to single layer graphene, without deteriorating the spatial resolution. The findings here point to improvement of graphene based nanopore devices for exciting opportunities in future single-molecule genomic screening devices.

Delayed dislocation of the radial head associated with malunion of distal radial fracture: A case report.

BACKGROUND: Traumatic radial head dislocation (RHD) is a well-described injury in the pediatric population. It is usually associated with an injury to the ulna in Monteggia fracture-dislocation, although it can occur as an isolated injury. Traumatic RHD with ipsilateral radial shaft fracture has rarely been reported. Delayed RHD secondary to the malunion of an isolated radial shaft fracture is extremely rare. CASE SUMMARY: We report a 9-year-old boy with limited pronation of the right elbow. The patient was diagnosed with delayed RHD associated with the malunion of a distal radial fracture. Since the annular ligament was disrupted with forearm rotation causing subluxation of the radial head, a modified double-strip Bell Tawse procedure was performed to reconstruct the annular ligament without corrective osteotomy for the malunited site. Four years after surgery, the angulation deformity of the distal radius was corrected with the restoration of the normal curvature of the radius. There was no recurrence of RHD. CONCLUSION: Annular ligament reconstruction without corrective osteotomy could reduce RHD and restore the normal curve of the radial shaft in children with delayed dislocation of the radial head associated with malunion of the radial shaft. Annular reconstruction using double triceps tendon strips might be useful for maintaining a more stable reduction by augmenting anterolateral parts.

Clinical Outcome after Clavicular Hook Plate Fixation for Displaced Medial-End Clavicle Fractures.

Background: Surgery of the medial end of the clavicle remains a challenge for orthopedic surgeons. Moreover, there is no standard surgical procedure for treating displaced fractures or dislocation of the medial clavicle. Thus, the present study aimed to evaluate the safety and efficacy of using a hook plate for treating medial-end clavicular fractures and present functional outcomes. Methods: We retrospectively investigated 18 patients who underwent surgery with a hook plate from July 2016 to December 2021. There were 14 men and 4 women with a mean age of 57.4 years. Fracture union was assessed at follow-up by computed tomography (CT). Other outcome parameters were complications, including implant failure, infection, nonunion, osteolysis of sternal manubrium, and migration of the hook portion. Range of motion (ROM), visual analog scale (VAS), Quick Disabilities of the Arm, Shoulder and Hand (Quick DASH), and American Shoulder and Elbow Society (ASES) scores were evaluated 6 months postoperatively and at the last follow-up. Results: The mean operation time was 43.8 minutes (range, 35-50 minutes) and the mean follow-up was 22.8 months (range, 12-42 months). Bone union was confirmed in all cases. The mean union time was 6.2 months (range, 6-7 months). Implant removal was performed routinely according to the clinical course in 17 cases. The mean implant removal time was 10.0 months (range, 6-14 months). Clinical and functional outcomes measured at the last follow-up were significantly improved compared to those at 6 months postoperatively (p < 0.05). Regarding complications, there were 6 cases (33.3%) of osteolysis of the sternal manubrium. Although the anteroposterior length of the manubrium and hook depth showed significant differences between the non-osteolysis group and the osteolysis group (p = 0.024), ROM, VAS, Quick DASH, and ASES scores were not significantly different between the two groups (all p > 0.05). Conclusions: Clavicle hook plating can be a safe and effective method that can be easily applied with good outcomes if it is used with appropriate surgical planning and technique for medial-end clavicle fracture. CT scans are useful for preoperative planning and postoperative evaluation of bone union or complications.

Breast Tissue Reconstruction Using Polycaprolactone Ball Scaffolds in a Partial Mastectomy Pig Model.

BACKGROUND: Breast cancer patients suffer from lowered quality of life (QoL) after surgery. Breast conservancy surgery (BCS) such as partial mastectomy is being practiced and studied as an alternative to solve this problem. This study confirmed breast tissue reconstruction in a pig model by fabricating a 3-dimensional (3D) printed Polycaprolactone spherical scaffold (PCL ball) to fit the tissue resected after partial mastectomy. METHODS: A 3D printed Polycaprolactone spherical scaffold with a structure that can help adipose tissue regeneration was produced using computer-aided design (CAD). A physical property test was conducted for optimization. In order to enhance biocompatibility, collagen coating was applied and a comparative study was conducted for 3 months in a partial mastectomy pig model. RESULTS: In order to identify adipose tissue and fibroglandular tissue, which mainly constitute breast tissue, the degree of adipose tissue and collagen regeneration was confirmed in a pig model after 3 months. As a result, it was confirmed that a lot of adipose tissue was regenerated in the PCL ball, whereas more collagen was regenerated in the collagen-coated Polycaprolactone spherical scaffold (PCL-COL ball). In addition, as a result of confirming the expression levels of TNF-a and IL-6, it was confirmed that PCL ball showed higher levels than PCL-COL ball. CONCLUSION: Through this study, we were able to confirm the regeneration of adipose tissue through a 3-dimensional structure in a pig model. Studies were conducted on medium and large-sized animal models for the final purpose of clinical use and reconstruction of human breast tissue, and the possibility was confirmed.

Cell-surface receptor for complement component C1q (gC1qR) is a key regulator for lamellipodia formation and cancer metastasis.

We previously demonstrated that the receptor for the complement component C1q (gC1qR) is a lipid raft protein that is indispensable for adipogenesis and insulin signaling. Here, we provide the first report that gC1qR is an essential component of lamellipodia in human lung carcinoma A549 cells. Cell-surface gC1qR was concentrated in the lamellipodia along with CD44, monosialoganglioside, actin, and phosphorylated focal adhesion kinase in cells stimulated with insulin, IGF-1, EGF, or serum. The growth factor-induced lamellipodia formation and cell migration were significantly decreased in gC1qR-depleted cells, with a concomitant blunt activation of the focal adhesion kinase and the respective receptor tyrosine kinases. Moreover, the gC1qR-depleted cells exhibited a reduced proliferation rate in culture as well as diminished tumorigenic and metastatic activities in grafted mice. We therefore conclude that cell-surface gC1qR regulates lamellipodia formation and metastasis via receptor tyrosine kinase activation.