Search NASA⌕ Search

SEARCH · Search NASA

Results for “NPC”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 73 records · Page 4
All sourcesNASA NTRSDOE OSTI
DOE OSTI2014

Integrative Structure–Function Mapping of the Nucleoporin Nup133 Suggests a Conserved Mechanism for Membrane Anchoring of the Nuclear Pore Complex

The nuclear pore complex (NPC) is the sole passageway for the transport of macromolecules across the nuclear envelope. Nup133, a major component in the essential Y-shaped Nup84 complex, is a large scaffold protein of the NPC's outer ring structure. Here, we describe an integrative modeling approach that produces atomic models for multiple states of Saccharomyces cerevisiae (Sc) Nup133, based on the crystal structures of the sequence segments and their homologs, including the related Vanderwaltozyma polyspora (Vp) Nup133 residues 55 to 502 (VpNup133 55–502 ) determined in this study, small angle X-ray scattering profiles for 18 constructs of ScNup133 and one construct of VpNup133, and 23 negative-stain electron microscopy class averages of ScNup1332–1157. Using our integrative approach, we then computed a multi-state structural model of the full-length ScNup133 and validated it with mutational studies and 45 chemical cross-links determined via mass spectrometry. Finally, the model of ScNup133 allowed us to annotate a potential ArfGAP1 lipid packing sensor (ALPS) motif in Sc and VpNup133 and discuss its potential significance in the context of the whole NPC; we suggest that ALPS motifs are scattered throughout the NPC's scaffold in all eukaryotes and play a major role in the assembly and membrane anchoring of the NPC in the nuclear envelope. Our results are consistent with a common evolutionary origin of Nup133 with membrane coating complexes (the protocoatomer hypothesis); the presence of the ALPS motifs in coatomer-like nucleoporins suggests an ancestral mechanism for membrane recognition present in early membrane coating complexes.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2021

Random sub-diffusion and capture of genes by the nuclear pore reduces dynamics and coordinates inter-chromosomal movement

Hundreds of genes interact with the yeast nuclear pore complex (NPC), localizing at the nuclear periphery and clustering with co-regulated genes. Dynamic tracking of peripheral genes shows that they cycle on and off the NPC and that interaction with the NPC slows their sub-diffusive movement. Furthermore, NPC-dependent inter-chromosomal clustering leads to coordinated movement of pairs of loci separated by hundreds of nanometers. We developed fractional Brownian motion simulations for chromosomal loci in the nucleoplasm and interacting with NPCs. These simulations predict the rate and nature of random sub-diffusion during repositioning from nucleoplasm to periphery and match measurements from two different experimental models, arguing that recruitment to the nuclear periphery is due to random sub-diffusion and transient capture by NPCs. Finally, the simulations do not lead to inter-chromosomal clustering or coordinated movement, suggesting that interaction with the NPC is necessary, but not sufficient, to cause clustering.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2018

PINK1 protects against oxidative stress induced senescence of human nucleus pulposus cells via regulating mitophagy

Intervertebral disc degeneration (IDD) is closely related with aging, whereas mitochondrial dysfunction is a common feature of aging in which results cell senescence. Phosphatase and tensin homolog (PTEN)-induced putative kinase protein 1 (PINK1) is a mitochondrial-targeted serine/threonine kinase, which plays a protective role against mitochondrial dysfunction with mitochondrial quality control by activating PINK1/Parkin mediated mitophagy. This study aimed to investigate the protective role of PINK1 against mitochondrial dysfunction and human nucleus pulposus cell (NPC) senescence. We found that mitochondrial dysfunction and NPC senescence could be induced under sublethal oxidative stress by 150 μM H{sub 2}O{sub 2}. Moreover, down-regulation of PINK1 tended to aggravate NPC senescence under oxidative stress. Therefore, mitophagy was evaluated in NPCs to further reveal the underlying mechanism. Results showed that sublethal oxidative stress induced mitochondria dysfunction and mitophagy in NPCs. Furthermore, depletion of PINK1 utilizing short hairpin RNA targeting PINK1 (PINK1-shRNA) impaired mitophagy, and exasperated NPC senescence under oxidative stress. In summary, these results suggested that PINK1 played as a protective role in clearance of damaged mitochondrial and alleviating cell senescence under oxidative stress, whose mechanism is associated with regulating mitophagy. These findings may provide a better understanding in pathomechanism of IDD and potential therapeutic approaches for IDD treatment.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

ABT-199-mediated inhibition of Bcl-2 as a potential therapeutic strategy for nasopharyngeal carcinoma

Highlights: • ABT-199 can effectively induce nasopharyngeal carcinoma cell apoptosis free of cytotoxic agent. • ABT-199 can cause elevated Noxa expression in nasopharyngeal carcinoma. • Dual Mcl-1/Bcl-2 inhibition with ABT-199 and Mcl-1 inhibitor S63845 works synergistically and demonstrates promise for future NPC treatment. Aberrant overexpression of Bcl-2 protein has been detected in 80% of nasopharyngeal carcinoma (NPC), and Bcl-2 family proteins are implicated in both NPC oncogenesis and chemotherapy resistance. Previous studies have shown that while treatment of NPC cells with Bcl-2 family inhibitors alone is rarely effective, concomitant treatment with a cytotoxic reagent such as cisplatin can increase efficacy through a synergistic effect. The aim of the current work was to determine how we might increase the efficacy of Bcl-2 family inhibitors in the absence of cytotoxic reagents, which are associated with negative side effect profiles.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Epstein-Barr virus Rta promotes invasion of bystander tumor cells through paracrine of matrix metalloproteinase 9

Highlights: • Rta-expressing cells promote invasion of bystander tumor cells through paracrine. • Rta upregulates MMP9 expression through activation of E2F. • The Rta-induced, paracrine-mediated pro-invasive effect is blocked upon knockdown or inhibition of MMP9. Clinical studies suggest a positive association between malignant progression of nasopharyngeal carcinoma (NPC) and Rta, a transcription factor of Epstein-Barr virus (EBV). However, Rta induces cellular senescence in vitro. To provide an underlying mechanism integrating these clues, we adapted a concept of senescence-associated secretory phenotype (SASP), based on which senescent cells facilitate tumor progression through paracrine. First, Rta-expressing NPC cells themselves show reduced invasiveness but promote invasion of Rta-negative tumor cells through secreted factors. Secretion of matrix metalloproteinase 9 (MMP9), an SASP protein, is increased by Rta, which requires the C-terminus of Rta and Rta-induced activation of E2F. Furthermore, the Rta-induced, paracrine-mediated pro-invasive effect is blocked upon knockdown of MMP9 expression or treatment with an MMP9 inhibitor. This study not only indicates that Rta can contribute to NPC progression through paracrine but also supports that MMP9 is a potential therapeutic target to prevent NPC metastasis.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2019

Achieving superior electromagnetic wave absorbers through the novel metal-organic frameworks derived magnetic porous carbon nanorods

High absorption capacity and broad absorption bandwidth electromagnetic wave (EMW) absorption materials (namely, EMW absorbers) are highly desirable due to the interference with electronics and harms on human beings’ health. In search for rational design on nanostructured absorbers, we have synthesized and demonstrated the rod-shape composites with Fe-containing magnetic nanoparticles (Fe 3 O 4 , Fe 3 C and Fe NPs) embedded into nano-porous carbon (NPC) through pyrolysis of Fe-based metal-organic frameworks (MOFs). The morphologies, compositions, and graphitization degree of the Fe-MOFs derived magnetic NPC nanorods can be effectively controlled via adjusting the pyrolysis temperatures. The graphitization level has a significant influence on the permittivity of the composites upon variation of pyrolysis temperatures, thereby a tunable electromagnetic wave (EMW) absorption is observed. Consequently, the resulting magnetic NPC nanorods obtained at pyrolysis temperature of 600 and 700 °C exhibit the most remarkable EMW absorption performance with a strong reflection loss of -52.9 dB and broad effective bandwidth (f e ) of 4.64 GHz at 3.07 mm. With a thickness of 3.5 mm, the f e for the magnetic NPC nanorods at 600 °C covers the whole X-band from 7.92 to 12.48 GHz. Finally, the noticeable EMW absorption performances have been greatly enhanced compared to those reported Fe 3 O 4 based absorbers, owing to the synergy of multiple components and the porous structures inherited from MOFs.

36 MATERIALS SCIENCE↗
DOE OSTI2012

Prognostic Value of Prevertebral Space Involvement in Nasopharyngeal Carcinoma Based on Intensity-Modulated Radiotherapy

Purpose: To investigate the prognostic significance of prevertebral space involvement (PSI) in patients with nasopharyngeal carcinoma (NPC) treated with intensity-modulated radiotherapy (IMRT). Methods and Materials: A retrospective review of data from 506 biopsy-proven, nonmetastatic NPCs was performed. Patients underwent magnetic resonance imaging examinations and received IMRT as their primary treatment. Results: In this series, 161 NPC patients (31.8%) had PSI. Parapharyngeal space (p < 0.001), skull base (p < 0.001), and paranasal sinuses (p = 0.009) were associated with PSI after multivariate analysis. The 4-year overall survival (OS), local relapse-free survival (LRFS), distant metastasis-free survival (DMFS) for NPC patients with and without PSI was 69.1% and 89.2% (p < 0.0001), 83.9% and 96.4% (p < 0.0001), and 71.6% and 89.6% (p < 0.0001), respectively. Multivariate analysis identified PSI as an independent negative prognostic factor for both OS (HR = 1.478-4.380; p = 0.001) and DMFS (HR = 1.389-4.174; p = 0.002). Patients with PSI had similar survival rates in OS and DMFS (p = 0.241 and p = 0.493, respectively) to that of T4 disease, while the differences between PSI and T3 disease in both OS and DMFS were distinctly significant (p = 0.029 and p = 0.029, respectively). Conclusions: For NPC patients treated with IMRT, PSI was found to be an independent prognostic factor for both OS and DMFS. It seems reasonable that PSI should be classified as a T4 disease on the basis of the current American Joint Committee on Cancer staging classification criteria.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2012

Intensity-Modulated Radiation Therapy in the Salvage of Locally Recurrent Nasopharyngeal Carcinoma

Purpose: Local recurrences of nasopharyngeal carcinoma (NPC) may be salvaged by reirradiation with conventional techniques, but with significant morbidity. Intensity-modulated radiation therapy (IMRT) may improve the therapeutic ratio by reducing doses to normal tissue. The aim of this study was to address the efficacy and toxicity profile of IMRT for a cohort of patients with locally recurrent NPC. Methods and Materials: Between August 2003 and June 2009, 70 patients with radiologic or pathologically proven locally recurrent NPC were treated with IMRT. The median time to recurrence was 30 months after the completion of conventional radiation to definitive dose. Fifty-seven percent of the tumors were classified asrT3-4. The minimum planned doses were 59.4 to 60 Gy in 1.8- to 2-Gy fractions per day to the gross disease with margins, with or without chemotherapy. Results: The median dose to the recurrent tumor was 70 Gy (range, 50-77.4 Gy). Sixty-five patients received the planned radiation therapy; 5 patients received between 50 and 60 Gy because of acute side effects. With a median follow-up time of 25 months, the rates of 2-year locoregional recurrence-free survival, disease-free survival, and overall survival were 65.8%, 65.8%, and 67.4%, respectively. Moderate to severe late toxicities were noted in 25 patients (35.7%). Eleven patients (15.7%) had posterior nasal space ulceration, 17 (24.3%) experienced cranial nerve palsies, 12 (17.1%) had trismus, and 12 (17.1%) experienced deafness. Extended disease-free interval (relative risk 2.049) and advanced T classification (relative risk 3.895) at presentation were adverse prognostic factors. Conclusion: Reirradiation with IMRT provides reasonable long-term control in patients with locally recurrent NPC.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2013

[{sup 18}F]fluoromisonidazole and a New PET System With Semiconductor Detectors and a Depth of Interaction System for Intensity Modulated Radiation Therapy for Nasopharyngeal Cancer

Purpose: The impact of a new type of positron emission tomography (New PET) with semiconductor detectors using {sup 18}F-labeled fluoromisonidazole (FMISO)-guided intensity modulated radiation therapy (IMRT) was compared with a state-of-the-art PET/computed tomography (PET/CT) system in nasopharyngeal cancer (NPC) patients. Methods and Materials: Twenty-four patients with non-NPC malignant tumors (control group) and 16 patients with NPC were subjected to FMISO-PET. The threshold of the tumor-to-muscle (T/M) ratio in each PET scan was calculated. The hypoxic volume within the gross tumor volume (GTVh) was determined using each PET ({sub NewPET}GTVh and {sub PET/CT}GTVh, respectively). Dose escalation IMRT plans prescribing 84 Gy to each GTVh were carried out. Results: The threshold of the T/M ratio was 1.35 for New PET and 1.23 for PET/CT. The mean volume of {sub NewPET}GTVh was significantly smaller than that of {sub PET/CT}GTVh (1.5 {+-} 1.6 cc vs 4.7 {+-} 4.6 cc, respectively; P=.0020). The dose escalation IMRT plans using New PET were superior in dose distribution to those using PET/CT. Dose escalation was possible in all 10 New PET-guided plans but not in 1 PET/CT-guided plan, because the threshold dose to the brainstem was exceeded. Conclusions: New PET was found to be useful for accurate dose escalation in FMISO-guided IMRT for patients with NPC.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2015

High-Sensitivity C-Reactive Protein Complements Plasma Epstein-Barr Virus Deoxyribonucleic Acid Prognostication in Nasopharyngeal Carcinoma: A Large-Scale Retrospective and Prospective Cohort Study

Purpose: To evaluate the effects of combining the assessment of circulating high-sensitivity C-reactive protein (hs-CRP) with that of Epstein-Barr virus DNA (EBV DNA) in the pretherapy prognostication of nasopharyngeal carcinoma (NPC). Patients and Methods: Three independent cohorts of NPC patients (training set of n=3113, internal validation set of n=1556, and prospective validation set of n=1668) were studied. Determinants of disease-free survival, distant metastasis–free survival, and overall survival were assessed by multivariate analysis. Hazard ratios and survival probabilities of the patient groups, segregated by clinical stage (T1-2N0-1M0, T3-4N0-1M0, T1-2N2-3M0, and T3-4N2-3M0) and EBV DNA load (low or high) alone, and also according to hs-CRP level (low or high), were compared. Results: Elevated hs-CRP and EBV DNA levels were significantly correlated with poor disease-free survival, distant metastasis–free survival, and overall survival in both the training and validation sets. Associations were similar and remained significant after excluding patients with cardiovascular disease, diabetes, and chronic hepatitis B. Patients with advanced-stage disease were segregated by high EBV DNA levels and high hs-CRP level into a poorest-risk group, and participants with either high EBV DNA but low hs-CRP level or high hs-CRP but low EBV DNA values had poorer survival compared with the bottom values for both biomarkers. These findings demonstrate a significant improvement in the prognostic ability of conventional advanced NPC staging. Conclusion: Baseline plasma EBV DNA and serum hs-CRP levels were significantly correlated with survival in NPC patients. The combined interpretation of EBV DNA with hs-CRP levels led to refinement of the risks for the patient subsets, with improved risk discrimination in patients with advanced-stage disease.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2016

Temporal Cerebral Microbleeds Are Associated With Radiation Necrosis and Cognitive Dysfunction in Patients Treated for Nasopharyngeal Carcinoma

Purpose: Radiation therapy for patients with nasopharyngeal carcinoma (NPC) may be complicated with radiation-induced brain necrosis (RN), resulting in deteriorated cognitive function. However, the underlying mechanism of this phenomenon remains unclear. This study attempts to elucidate the association between cerebral microbleeds (CMBs) and radiation necrosis and cognitive dysfunction in NPC patients treated with radiation therapy. Methods and Materials: This cross-sectional study included 106 NPC patients who were exposed to radiation therapy (78 patients with RN and 28 without RN). Sixty-six patients without discernable intracranial pathology were included as the control group. CMBs were confirmed using susceptibility-weighted magnetic resonance imaging. Cognitive function was accessed using Montreal Cognitive Assessment. Patients with a total score below 26 were defined as cognitively dysfunction. Results: Seventy-seven patients (98.7%) in the RN group and 12 patients (42.9%) in the non-RN group had at least 1 CMB. In contrast, only 14 patients (21.2%) in the control group had CMBs. In patients with a history of radiation therapy, CMBs most commonly presented in temporal lobes (76.4%) followed by cerebellum (23.7%). Patients with RN had more temporal CMBs than those in the non-RN group (37.7 ± 51.9 vs 3.8 ± 12.6, respectively; P<.001). The number of temporal lobe CMBs was predictive for larger volume of brain necrosis (P<.001) in multivariate linear regression analysis. Although cognitive impairment was diagnosed in 55.1% of RN patients, only 7.1% of non-RN patients sustained cognitive impairment (P<.001). After adjusting for age, sex, education, period after radiation therapy, CMBs in other lobes, and RN volume, the number of temporal CMBs remained an independent risk factor for cognitive dysfunction (odds ratio [OR]: 1.03; 95% confidence interval [CI]: 1.01-1.04; P=.003). Conclusions: CMBs is a common radiological manifestation in NPC patients with RN. The number of temporal CMBs is independently associated with increased likelihood of cognitive dysfunction in patients with RN.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2016

Comparison of Planning Quality and Efficiency Between Conventional and Knowledge-based Algorithms in Nasopharyngeal Cancer Patients Using Intensity Modulated Radiation Therapy

Purpose: Intensity modulated radiation therapy (IMRT) is widely used to achieve a highly conformal dose and improve treatment outcome. However, plan quality and planning time are institute and planner dependent, and no standardized tool exists to recognize an optimal plan. RapidPlan, a knowledge-based algorithm, can generate constraints to assist optimization and produce high-quality IMRT plans. This report evaluated the quality and efficiency of using RapidPlan in nasopharyngeal carcinoma (NPC) IMRT planning. Methods and Materials: RapidPlan was configured using 79 radical IMRT plans for NPC; 20 consecutive NPC patients indicated for radical radiation therapy between October 2014 and May 2015 were then recruited to assess its performance. The ability of RapidPlan to produce acceptable plans was evaluated. For plans that could not achieve clinical acceptance, manual touch-up was performed. The IMRT plans produced without RapidPlan (manual plans) and with RapidPlan (RP-2 plans, including those with manual touch-up) were compared in terms of dosimetric quality and planning efficiency. Results: RapidPlan by itself could produce clinically acceptable plans for 9 of the 20 patients; manual touch-up increased the number of acceptable plans (RP-2 plans) to 19. The target dose coverage and conformity were very similar. No difference was found in the maximum dose to the brainstem and optic chiasm. RP-2 plans delivered a higher maximum dose to the spinal cord (46.4 Gy vs 43.9 Gy, P=.002) but a lower dose to the parotid (mean dose to right parotid, 37.3 Gy vs 45.4 Gy; left, 34.4 Gy vs 43.1 Gy; P<.001) and the right cochlea (mean dose, 48.6 Gy vs 52.6 Gy; P=.02). The total planning time for RP-2 plans was significantly less than that for manual plans (64 minutes vs 295 minutes, P<.001). Conclusions: This study shows that RapidPlan can significantly improve planning efficiency and produce quality IMRT plans for NPC patients.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2017

Reporting Quality of Randomized, Controlled Trials Evaluating Combined Chemoradiotherapy in Nasopharyngeal Carcinoma

Purpose: To comprehensively assess the reporting quality of randomized, controlled trials (RCTs) in nasopharyngeal carcinoma (NPC), and to identify significant predictors of quality. Methods and Materials: Two investigators searched MEDLINE and EMBASE for RCTs published between January 1988 and December 2015 that assessed the effect of combined chemoradiotherapy for NPC. The overall quality of each report was assessed using a 28-point overall quality score (OQS) based on the 2010 Consolidated Standards of Reporting Trials (CONSORT) statement. To provide baseline data for further evaluation, we also investigated the reporting quality of certain important issues in detail, including key methodologic items (allocation concealment, blinding, intention-to-treat principle), endpoints, follow-up, subgroup analyses, and adverse events. Results: We retrieved 24 relevant RCTs including 6591 patients. Median 2010 OQS was 15.5 (range, 10-24). Half of the items in the 2010 OQS were poorly reported in at least 40% of trials. Multivariable regression models revealed that publication after 2010 and high impact factor were significant predictors of improved 2010 OQS. Additionally, many issues that we consider significant were not reported adequately. Conclusions: Despite publication of the CONSORT statement more than a decade ago, overall reporting quality for RCTs in NPC was unsatisfactory. Additionally, substantial selectivity and heterogeneity exists in reporting of certain crucial issues. This survey provides the first prompt for NPC trial investigators to improve reporting quality according to the CONSORT statement; increased scrutiny and diligence by editors and peer reviewers is also required.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2017

Evaluation of deformed image-based dose calculations for adaptive radiotherapy of nasopharyngeal carcinoma

The ultimate goal of adaptive radiotherapy (ART) is to deliver truly customized radiation treatments. Currently, the quality of cone-beam computed tomography (CBCT) images is still inferior to that of conventional CT images in contour delineations and dose calculations for replanning purposes. This retrospective study aims to evaluate the dosimetric accuracy of using deformed conventional CT images for dose calculations, in the hope of inferring the feasibility of ART using planning CT (PCT) images that deformed to up-to-date CBCT images for patients with nasopharyngeal carcinoma (NPC). Thirty consecutive patients with NPC who had undergone 1 replan in their radiotherapy treatments were selected. The pretreatment PCT images were deformed to match the mid-treatment PCT images by deformable image registration. The same volumetric modulated arc therapy plan was then calculated on the deformed PCT images. The resulting dose distributions and dose volume histograms of the tumors and organs at risk (OARs) were compared with the original plan. Five dose levels, D{sub 98%}, D{sub 95%}, D{sub 50%}, D{sub 5%}, and D{sub 2%}, were recorded for 9 NPC targets. Four dose levels, D{sub max}, D{sub 10%}, D{sub 50%}, and D{sub mean}, were recorded for 15 OARs. The greatest percentage difference in observed dose for D{sub 98%}, D{sub 95%}, D{sub 50%}, D{sub 5%}, and D{sub 2%} of the targets were 1.71%, 1.55%, 0.64%, 0.97%, and 1.13%, respectively. The greatest percentage difference in observed dose for D{sub max}, D{sub 10%}, D{sub 50%}, and D{sub mean} of the OARs were -26.51% (left optic nerve), -17.06% (left optic nerve), 56.70% (spinal cord), and 18.97% (spinal cord), respectively. In addition, 29 of 45 (64%) dosimetric end points of the targets showed statistically significant dose differences (p p < 0.05). Dose calculations using deformed PCT images could result in significant dose uncertainties in target volumes and OARs. Larger dose deviations were found in OARs in comparison with target volumes. The spinal cord and optic nerve showed the greatest percentage dose differences and the clinical significance has yet to be determined. Deformable registration error was believed to be the problem causing the dose deviations. Owing to unknown clinical significanceof dose deviation results obtained from this study, a conventional CT scan is still required for replanning in patients with NPC who are experiencing significant anatomical changes during the course of radiation treatment.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
DOE OSTI2018

The formation of giant plasma membrane vesicles enable new insights into the regulation of cholesterol efflux

Highlights: • Giant plasma membrane vesicles serve as a means to monitor cell surface cholesterol. • Increase in cholesterol efflux enhances giant plasma membrane vesicle formation. • Microtubule stabilization and acetylation promote cholesterol efflux in NPC disease. • Actin dynamics and Arl4c modulate lipid efflux and giant plasma membrane vesicles. Aberrant cellular cholesterol accumulation contributes to the pathophysiology of many diseases including neurodegenerative disorders such as Niemann-Pick Type C (NPC) and Alzheimer's Disease{sup 1–4}. Many aspects of cholesterol efflux from cells remain elusive. Here we describe the utility of cholesterol-rich giant plasma membrane vesicles (GPMVs) as a means to monitor cholesterol that is translocated to the plasma membrane for secretion. We demonstrate that small molecules known to enhance lipid efflux, including those in clinical trials for lipid storage disorders, enhance this GPMV formation. Conversely, pharmacological inhibition of cholesterol efflux blocks GPMV formation. We show that microtubule stabilization via paclitaxel treatment and increased tubulin acetylation via HDAC6 inhibition promotes the formation of GPMVs with concomitant reduction in cellular cholesterol in a cell model of NPC disease. The pan-deacetylase inhibitor panobinostat, which has been shown to reduce the severity of cholesterol storage in NPC, elicited a similar response. Further, the disruption of actin polymerization inhibits the formation of GPMVs, whereas the small GTP-binding protein Arl4c promotes actin remodeling at sites overlapping with GPMV formation. Thus, monitoring the formation of GPMVs provides a new avenue to better understand diseases whose pathology may be sensitive to alterations in cellular cholesterol.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2022

Structure of cytoplasmic ring of nuclear pore complex by integrative cryo-EM and AlphaFold

The nuclear pore complex (NPC) is the conduit for bidirectional cargo traffic between the cytoplasm and the nucleus. We determined a near-complete structure of the cytoplasmic ring of the NPC from Xenopus oocytes using single-particle cryo–electron microscopy and AlphaFold prediction. Structures of nucleoporins were predicted with AlphaFold and fit into the medium-resolution map by using the prominent secondary structural density as a guide. Certain molecular interactions were further built or confirmed by complex prediction by using AlphaFold. Here, we identified the binding modes of five copies of Nup358, the largest NPC subunit with Phe-Gly repeats for cargo transport, and predicted it to contain a coiled-coil domain that may provide avidity to assist its role as a nucleation center for NPC formation under certain conditions

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Role of pore dilation in molecular transport through the nuclear pore complex: Insights from polymer scaling theory

The nuclear pore complex (NPC), a channel within the nuclear envelope filled with intrinsically disordered proteins, regulates the transport of macromolecules between the nucleus and the cytoplasm. Recent studies have highlighted the NPC’s ability to adjust its diameter in response to the membrane tension, underscoring the importance of exploring how variations in pore size influence molecular transport through the NPC. In this study, we investigated the relationship between pore size and transport rate and proposed a mathematical model describing this connection. We began by theoretically analyzing how the pore size scales with the characteristic dimensions of the mesh-like structure within the pore. By introducing key assumptions about how the meshwork structure influences molecular diffusion, we derived a mathematical expression for the transport rate based on the size of the pore and the transported molecules. To validate our model, we conducted Brownian dynamics simulations using a coarse-grained representation of the NPC. These simulations, performed across a range of pore sizes, demonstrated strong agreement with our model’s predictions, confirming its accuracy and applicability. Our model is specifically tailored for small-to-medium-sized molecules, approximately 5 nanometers in size, making it relevant to a wide range of transcription factors and signaling molecules. It also extends to molecules with weak and transient interactions with FG-Nups, such as importin-β. By presenting this model formula, our study offers a quantitative framework for analyzing the effects of pore dilation on nucleocytoplasmic transport.

Biochemistry & Molecular Biology↗