175 results on '"Weinberg MS"'
Search Results
2. MULTIPLE SETS OF HUMAN INDEXING FOR CIVIL ENGINEERING DOCUMENTS
- Author
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Dls Bella Hass Weinberg Ms
- Subjects
Set (abstract data type) ,Information retrieval ,Index (publishing) ,Computer science ,Automatic indexing ,As is ,Search engine indexing ,Library and Information Sciences ,Civil engineering ,Cross-reference ,Field (computer science) - Abstract
Four sets of indexing from various sources for sixty-five journal artcles in the field of civil engineering are analyzed statistically and linguistically. Cross reference patterns of the sets, which include Engineering Index and Applied Science and Technology Index, as well as relevant thesauri are examined. The percentage of terms from each set which are found in the authors' abstracts and full text is reported, as is the contribution of cross references to the find rate. The implicatsion of some of the unexpected findings for free-text online searching and for automatic indexing are given, and the methodological problems of comparing terms from different sources are discussed.
- Published
- 1982
3. Thiazide Induced Hypotension: The Role of Plasma Volume Reduction and the Urinary Kallikrein System
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Solomon Rj, Stillman N, and Weinberg Ms
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medicine.medical_specialty ,Mean arterial pressure ,business.industry ,Stimulation ,Plasma volume ,Essential hypertension ,medicine.disease ,Plasma renin activity ,Excretion ,Endocrinology ,Blood pressure ,Internal medicine ,medicine ,business ,Thiazide ,medicine.drug - Abstract
The hypotensive response the thiazide diuretics was studied in 15 males with moderate essential hypertension and correlated with serial changes in plasma volume, weight, plasma renin activity, urinary aldosterone, and urinary kallikrein, both total and activity. A greater than 10 mmHg fall in mean arterial pressure after four weeks of treatment defined the responders to therapy (n = 10) while all others were considered non-responders (n = 5). In responders, the fall in mean arterial pressure was accompanied by sustained reduction in plasma volume and weight. No sustained fall in plasma volume was noted in non-responders. Plasma renin activity and urinary aldosterone excretion increased in responders but not in non-responders. Urinary kallikrein, both total and active, increased in the responders but remained unchanged in the non-responders. The results are consistent with the hypothesis that a sustained reduction in plasma volume is necessary for the maintenance of a hypotensive response to thiazides. Stimulation of the renal kallikrein-kinin system may be necessary to balance the antinatriuretic and pressor effects of the renin-angiotensin-aldosterone system. If unopposed, this system would return plasma volume and blood pressure to pretreatment levels.
- Published
- 1986
4. Letter to the editor
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Weinberg Ms
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Clinical tests ,Text mining ,business.industry ,Computer science ,Artificial intelligence ,Toxicology ,computer.software_genre ,business ,computer ,Natural language processing - Published
- 1989
5. Long-term safety of high-dose angiotensin receptor blocker therapy in hypertensive patients with chronic kidney disease.
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Weinberg AJ, Zappe DH, Ramadugu R, and Weinberg MS
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- 2006
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6. Tuberculosis in United States-Bound Follow-to-Join Asylees, 2014-2019.
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Liu Y, Posey DL, Weinberg MS, and Phares CR
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- Humans, United States epidemiology, Female, Male, Adolescent, Young Adult, Child, Adult, Child, Preschool, Mass Screening methods, Tuberculosis diagnosis, Tuberculosis epidemiology, Refugees statistics & numerical data
- Abstract
Persons may seek asylum in the United States or at a U.S. port of entry. Principal asylees are those who are granted asylum status. Their spouse and unmarried children under 21 years of age may be granted asylum if accompanying, or following to join, the principal asylees. U.S.-bound follow-to-join asylees must undergo an overseas medical examination that includes tuberculosis (TB) screening. Culture-based overseas TB screening in U.S.-bound follow-to-join asylees has not been evaluated. We evaluated data from overseas TB screening in 19,088 arrivals of follow-to-join asylees during 2014-2019 and assessed data from their postarrival evaluation, which is recommended for those at risk for TB. Of 19,088 arrivals of follow-to-join asylees, 29 (152 cases/100,000 persons) met criteria for class B0 TB (recent completion of TB treatment overseas) and 340 (1,781 cases/100,000 persons) met criteria for class B1 pulmonary TB (chest radiograph/clinical symptoms suggestive of TB but negative sputum cultures overseas). Of 6,847 persons aged 2 to 14 years from countries with a WHO-estimated TB incidence of ≥20 cases/100,000 population/year, 408 (6.0%) were classified as class B2 latent TB infection (LTBI). Postarrival evaluations were completed in 44.8%, 51.5%, and 40.4% of persons with class B0 TB, class B1 TB, and class B2 LTBI, respectively. In conclusion, culture-based overseas TB screening in U.S.-bound follow-to-join asylees is effective in identifying those with TB (class B0 TB) or those at risk for TB (class B1 TB and class B2 LTBI). Completion of postarrival evaluation for newly arrived follow-to-join asylees was less frequent than that reported for immigrants and refugees.
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- 2024
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7. Cryopreservation of cerebrospinal fluid cells preserves the transcriptional landscape for single-cell analysis.
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Kodali MC, Antone J, Alsop E, Jayakumar R, Parikh K, Chiot A, Sanchez-Molina P, Ajami B, Arnold SE, Jensen K, Das S, and Weinberg MS
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- Humans, Cells, Cultured, Cryopreservation methods, Single-Cell Analysis, Cell Survival, Dimethyl Sulfoxide pharmacology, Cryoprotective Agents pharmacology
- Abstract
Cerebrospinal fluid (CSF) matrix biomarkers have become increasingly valuable surrogate markers of neuropsychiatric diseases in research and clinical practice. In contrast, CSF cells have been rarely investigated due to their relative scarcity and fragility, and lack of common collection and cryopreservation protocols, with limited exceptions for neurooncology and primary immune-based diseases like multiple sclerosis. the advent of a microfluidics-based multi-omics approach to studying individual cells has allowed for the study of cellular phenotyping, intracellular dynamics, and intercellular relationships that provide multidimensionality unable to be obtained through acellular fluid-phase analyses. challenges to cell-based research include site-to-site differences in handling, storage, and thawing methods, which can lead to inaccuracy and inter-assay variability. In the present study, we performed single-cell RNA sequencing (10x Genomics) on fresh or previously cryopreserved human CSF samples from three alternative cryopreservation methods: Fetal Bovine Serum with Dimethyl sulfoxide (FBS/DMSO), FBS/DMSO after a DNase step (a step often included in epigenetic studies), and cryopreservation using commercially available Recovery© media. In comparing relative differences between fresh and cryopreserved samples, we found little effect of the cryopreservation method on being able to resolve donor-linked cell type proportions, markers of cellular stress, and overall gene expression at the single-cell level, whereas donor-specific differences were readily discernable. We further demonstrate the compatibility of fresh and cryopreserved CSF immune cell sequencing using biologically relevant sexually dimorphic gene expression differences by donor. Our findings support the utility and interchangeability of FBS/DMSO and Recovery cryopreservation with fresh sample analysis, providing a methodological grounding that will enable researchers to further expand our understanding of the CSF immune cell contributions to neurological and psychiatric disease., (© 2024. The Author(s).)
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- 2024
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8. Effect of Metformin on Plasma and Cerebrospinal Fluid Biomarkers in Non-Diabetic Older Adults with Mild Cognitive Impairment Related to Alzheimer's Disease.
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Weinberg MS, He Y, Kivisäkk P, Arnold SE, and Das S
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- Humans, Aged, Male, Female, Proteomics methods, Double-Blind Method, Aged, 80 and over, Middle Aged, Metformin therapeutic use, Alzheimer Disease cerebrospinal fluid, Alzheimer Disease blood, Alzheimer Disease drug therapy, Cognitive Dysfunction cerebrospinal fluid, Cognitive Dysfunction blood, Cognitive Dysfunction drug therapy, Biomarkers blood, Biomarkers cerebrospinal fluid, Hypoglycemic Agents therapeutic use
- Abstract
Background: Alzheimer's disease (AD) is a complicated condition involving multiple metabolic and immunologic pathophysiological processes that can occur with the hallmark pathologies of amyloid-β, tau, and neurodegeneration. Metformin, an anti-diabetes drug, targets several of these disease processes in in vitro and animal studies. However, the effects of metformin on human cerebrospinal fluid (CSF) and plasma proteins as potential biomarkers of treatment remain unexplored., Objective: Using proteomics data from a metformin clinical trial, identify the impact of metformin on plasma and CSF proteins., Methods: We analyzed plasma and CSF proteomics data collected previously (ClinicalTrials.gov identifier: NCT01965756, conducted between 2013 and 2015), and conduced bioinformatics analyses to compare the plasma and CSF protein levels after 8 weeks of metformin or placebo use to their baseline levels in 20 non-diabetic patients with mild cognitive impairment (MCI) and positive AD biomarkers participants., Results: 50 proteins were significantly (unadjusted p < 0.05) altered in plasma and 26 in CSF after 8 weeks of metformin use, with 7 proteins in common (AZU1, CASP-3, CCL11, CCL20, IL32, PRTN3, and REG1A). The correlation between changes in plasma and CSF levels of these 7 proteins after metformin use relative to baseline levels was high (r = 0.98). The proteins also demonstrated temporal stability., Conclusions: Our pilot study is the first to investigate the effect of metformin on plasma and CSF proteins in non-diabetic patients with MCI and positive AD biomarkers and identifies several candidate plasma biomarkers for future clinical trials after confirmatory studies.
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- 2024
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9. Screening, Assessment, and Pharmacologic Treatment of Mild Cognitive Impairment and Early Alzheimer's Disease: The Role for Monoclonal Antibodies.
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Weinberg MS, Principe JL, Chen A, Chung SY, Arnold SE, and Stern TA
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- Humans, Antibodies, Monoclonal, Hospitals, General, Referral and Consultation, Alzheimer Disease complications, Alzheimer Disease diagnosis, Alzheimer Disease drug therapy, Cognitive Dysfunction complications, Cognitive Dysfunction diagnosis, Cognitive Dysfunction drug therapy, Mental Disorders therapy, Psychiatry
- Abstract
The Psychiatric Consultation Service at Massachusetts General Hospital sees medical and surgical inpatients with comorbid psychiatric symptoms and conditions. During their twice-weekly rounds, Dr Stern and other members of the Consultation Service discuss diagnosis and management of hospitalized patients with complex medical or surgical problems who also demonstrate psychiatric symptoms or conditions. These discussions have given rise to rounds reports that will prove useful for clinicians practicing at the interface of medicine and psychiatry., Prim Care Companion CNS Disord. 2023;25(6):23f03544 ., Author affiliations are listed at the end of this article., (© Copyright 2023 Physicians Postgraduate Press, Inc.)
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- 2023
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10. Association of BCG Vaccine Treatment With Death and Dementia in Patients With Non-Muscle-Invasive Bladder Cancer.
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Weinberg MS, Zafar A, Magdamo C, Chung SY, Chou WH, Nayan M, Deodhar M, Frendl DM, Feldman AS, Faustman DL, Arnold SE, Vakulenko-Lagun B, and Das S
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- Male, Humans, Aged, Female, BCG Vaccine therapeutic use, Adjuvants, Immunologic, Cohort Studies, Administration, Intravesical, Non-Muscle Invasive Bladder Neoplasms, Urinary Bladder Neoplasms epidemiology, Urinary Bladder Neoplasms drug therapy, Dementia epidemiology, Dementia drug therapy
- Abstract
Importance: The BCG vaccine-used worldwide to prevent tuberculosis-confers multiple nonspecific beneficial effects, and intravesical BCG vaccine is currently the recommended treatment for non-muscle-invasive bladder cancer (NMIBC). Moreover, BCG vaccine has been hypothesized to reduce the risk of Alzheimer disease and related dementias (ADRD), but previous studies have been limited by sample size, study design, or analyses., Objective: To evaluate whether intravesical BCG vaccine exposure is associated with a decreased incidence of ADRD in a cohort of patients with NMIBC while accounting for death as a competing event., Design, Setting, and Participants: This cohort study was performed in patients aged 50 years or older initially diagnosed with NMIBC between May 28, 1987, and May 6, 2021, treated within the Mass General Brigham health care system. The study included a 15-year follow-up of individuals (BCG vaccine treated or controls) whose condition did not clinically progress to muscle-invasive cancer within 8 weeks and did not have an ADRD diagnosis within the first year after the NMIBC diagnosis. Data analysis was conducted from April 18, 2021, to March 28, 2023., Main Outcomes and Measures: The main outcome was time to ADRD onset identified using diagnosis codes and medications. Cause-specific hazard ratios (HRs) were estimated using Cox proportional hazards regression after adjusting for confounders (age, sex, and Charlson Comorbidity Index) using inverse probability scores weighting., Results: In this cohort study including 6467 individuals initially diagnosed with NMIBC between 1987 and 2021, 3388 patients underwent BCG vaccine treatment (mean [SD] age, 69.89 [9.28] years; 2605 [76.9%] men) and 3079 served as controls (mean [SD] age, 70.73 [10.00] years; 2176 [70.7%] men). Treatment with BCG vaccine was associated with a lower rate of ADRD (HR, 0.80; 95% CI, 0.69-0.99), with an even lower rate of ADRD in patients aged 70 years or older at the time of BCG vaccine treatment (HR, 0.74; 95% CI, 0.60-0.91). In competing risks analysis, BCG vaccine was associated with a lower risk of ADRD (5-year risk difference, -0.011; 95% CI, -0.019 to -0.003) and a decreased risk of death in patients without an earlier diagnosis of ADRD (5-year risk difference, -0.056; 95% CI, -0.075 to -0.037)., Conclusions and Relevance: In this study, BCG vaccine was associated with a significantly lower rate and risk of ADRD in a cohort of patients with bladder cancer when accounting for death as a competing event. However, the risk differences varied with time.
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- 2023
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11. Non-uniform dystrophin re-expression after CRISPR-mediated exon excision in the dystrophin/utrophin double-knockout mouse model of DMD.
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Hanson B, Stenler S, Ahlskog N, Chwalenia K, Svrzikapa N, Coenen-Stass AML, Weinberg MS, Wood MJA, and Roberts TC
- Abstract
Duchenne muscular dystrophy (DMD) is the most prevalent inherited myopathy affecting children, caused by genetic loss of the gene encoding the dystrophin protein. Here we have investigated the use of the Staphylococcus aureus CRISPR-Cas9 system and a double-cut strategy, delivered using a pair of adeno-associated virus serotype 9 (AAV9) vectors, for dystrophin restoration in the severely affected dystrophin/utrophin double-knockout (dKO) mouse. Single guide RNAs were designed to excise Dmd exon 23, with flanking intronic regions repaired by non-homologous end joining. Exon 23 deletion was confirmed at the DNA level by PCR and Sanger sequencing, and at the RNA level by RT-qPCR. Restoration of dystrophin protein expression was demonstrated by western blot and immunofluorescence staining in mice treated via either intraperitoneal or intravenous routes of delivery. Dystrophin restoration was most effective in the diaphragm, where a maximum of 5.7% of wild-type dystrophin expression was observed. CRISPR treatment was insufficient to extend lifespan in the dKO mouse, and dystrophin was expressed in a within-fiber patchy manner in skeletal muscle tissues. Further analysis revealed a plethora of non-productive DNA repair events, including AAV genome integration at the CRISPR cut sites. This study highlights potential challenges for the successful development of CRISPR therapies in the context of DMD., Competing Interests: The authors declare no competing interests., (© 2022 The Author(s).)
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- 2022
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12. Effectiveness-Essential for Cost-effectiveness-Reply.
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Ross EL, Weinberg MS, and Arnold SE
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- 2022
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13. Psychedelics for Alzheimer's Disease Palliative Care.
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McManus KR, Patrick R, Striepe MI, Drury MJ, Ozonsi R, Forester BP, and Weinberg MS
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- 2022
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14. Multidrug-Resistant Tuberculosis in U.S.-Bound Immigrants and Refugees.
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Liu Y, Posey DL, Yang Q, Weinberg MS, Maloney SA, Lambert LA, Ortega LS, Marano N, Cetron MS, and Phares CR
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- Antitubercular Agents pharmacology, Antitubercular Agents therapeutic use, Humans, Isoniazid pharmacology, Rifampin, United States epidemiology, Emigrants and Immigrants, Extensively Drug-Resistant Tuberculosis, Mycobacterium tuberculosis, Refugees, Tuberculosis, Multidrug-Resistant drug therapy, Tuberculosis, Multidrug-Resistant epidemiology
- Abstract
Rationale: Approximately two-thirds of new cases of tuberculosis (TB) in the United States are among non-U.S.-born persons. Culture-based overseas TB screening in U.S.-bound immigrants and refugees has substantially reduced the importation of TB into the United States, but it is unclear to what extent this program prevents the importation of multidrug-resistant TB (MDR-TB). Objectives: To study the epidemiology of MDR-TB in U.S.-bound immigrants and refugees and to evaluate the effect of culture-based overseas TB screening in U.S.-bound immigrants and refugees on reducing the importation of MDR-TB into the United States. Methods: We analyzed data of immigrants and refugees who completed overseas treatment for culture-positive TB during 2015-2019. We also compared mean annual number of MDR-TB cases in non-U.S.-born persons within 1 year of arrival in the United States between 1996-2006 (when overseas screening followed a smear-based algorithm) and 2014-2019 (after full implementation of a culture-based algorithm). Results: Of 3,300 culture-positive TB cases identified by culture-based overseas TB screening in immigrants and refugees during 2015-2019, 122 (3.7%; 95% confidence interval [CI], 3.1-4.1) had MDR-TB, 20 (0.6%; 95% CI, 0.3-0.9) had rifampicin-resistant TB, 382 (11.6%; 95% CI, 10.5-12.7) had isoniazid-resistant TB, and 2,776 (84.1%; 95% CI, 82.9-85.4) had rifampicin- and isoniazid-susceptible TB. None were diagnosed with extensively drug-resistant TB. All 3,300 persons with culture-positive TB completed treatment overseas; of 70 and 11 persons who were treated overseas for MDR-TB and rifampicin-resistant TB, respectively, none were diagnosed with TB disease at postarrival evaluation in the United States. Culture-based overseas TB screening in U.S.-bound immigrants and refugees prevented 24.4 MDR-TB cases per year from arriving in the United States, 18.2 cases more than smear-based overseas TB screening. The mean annual number of MDR-TB cases among non-U.S.-born persons within 1 year of arrival in the United States decreased from 34.6 cases in 1996-2006 to 19.5 cases in 2014-2019 (difference of 15.1; P < 0.001). Conclusions: Culture-based overseas TB screening in U.S.-bound immigrants and refugees substantially reduced the importation of MDR-TB into the United States.
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- 2022
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15. Cost-effectiveness of Aducanumab and Donanemab for Early Alzheimer Disease in the US.
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Ross EL, Weinberg MS, and Arnold SE
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- Adult, Aged, Amyloid, Antibodies, Monoclonal therapeutic use, Antibodies, Monoclonal, Humanized, Cost-Benefit Analysis, Disease Progression, Humans, Alzheimer Disease drug therapy
- Abstract
Importance: Several anti-amyloid monoclonal antibodies have been developed for slowing the progression of Alzheimer disease (AD). Among the furthest developed are aducanumab, which received accelerated approval from the US Food and Drug Administration in 2021, and donanemab, which is currently undergoing phase 3 trials. The cost-effectiveness of these treatments has not been established., Objectives: To estimate the cost-effectiveness of aducanumab and donanemab relative to standard care for early AD in the US., Design, Setting, and Participants: A decision analytic model was used to estimate the lifetime health and economic outcomes of adults with early AD, from US healthcare sector and societal perspectives. Simulated patients had a mean (SD) age of 75.2 (5.5) years; 65% had mild cognitive impairment and 35% had mild dementia. Analyses were conducted from April 6, 2021, to January 20, 2022., Interventions: Standard care, aducanumab (selected inputs including disease progression hazard ratio [HR] of 0.89 [95% CI, 0.63-1.15], annual price of $28 000, and twice-yearly monitoring with magnetic resonance imaging [MRI] of the brain), or donanemab (selected inputs including disease progression HR of 0.68 [95% CI, 0.44-0.99], annual price of $28 000, and twice-yearly monitoring with MRI of the brain and amyloid positron emission tomography [PET] monitoring). Donanemab was switched to placebo after substantial amyloid reduction on PET imaging, which occurred in 27% of patients at 6 months and 55% of patients at 12 months., Main Outcomes and Measures: Quality-adjusted life-years (QALYs); costs, in 2020 US dollars; incremental cost-effectiveness ratios (ICERs); and value-based prices, defined as the maximum price at which a treatment would be cost-effective given a cost-effectiveness threshold of ICER of $150 000/QALY., Results: Lifetime QALYs increased by 0.133 with aducanumab and 0.408 with donanemab. Total health care sector and societal costs increased by $130 100 and $127 800, respectively, with aducanumab and by $78 700 and $71 600, respectively, with donanemab, driven largely by drug costs ($119 000 for aducanumab and $44 600 for donanemab). Health care sector and societal ICERs relative to standard care were $981 000/QALY and $964 000/QALY, respectively, for aducanumab and $193 000/QALY and $176 000/QALY, respectively, for donanemab. In sensitivity analysis, aducanumab's value-based price remained less than $50 000/y, even when assuming a 90% reduction in disease progression. Donanemab's value-based price surpassed $50 000/y once its efficacy exceeded 50%., Conclusions and Relevance: These findings suggest that at current expected prices, neither aducanumab nor donanemab would be cost-effective for early AD in the US. Donanemab's dosing scheme, in which patients suspend treatment on achieving substantial amyloid reductions, may provide a rubric by which sufficiently effective anti-amyloid antibody treatments could be cost-effective even when priced comparably to other biologics.
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- 2022
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16. Nuclear microRNA-466c regulates Vegfa expression in response to hypoxia.
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Laitinen P, Väänänen MA, Kolari IL, Mäkinen PI, Kaikkonen MU, Weinberg MS, Morris KV, Korhonen P, Malm T, Ylä-Herttuala S, Roberts TC, Turunen MP, and Turunen TA
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- Animals, Hypoxia genetics, Mice, RNA, Messenger, MicroRNAs genetics, RNA, Long Noncoding genetics, Vascular Endothelial Growth Factor A metabolism
- Abstract
MicroRNAs are well characterized in their role in silencing gene expression by targeting 3´-UTR of mRNAs in cytoplasm. However, recent studies have shown that miRNAs have a role in the regulation of genes in the nucleus, where they are abundantly located. We show here that in mouse endothelial cell line (C166), nuclear microRNA miR-466c participates in the regulation of vascular endothelial growth factor a (Vegfa) gene expression in hypoxia. Upregulation of Vegfa expression in response to hypoxia was significantly compromised after removal of miR-466c with CRISPR-Cas9 genomic deletion. We identified a promoter-associated long non-coding RNA on mouse Vegfa promoter and show that miR-466c directly binds to this transcript to modulate Vegfa expression. Collectively, these observations suggest that miR-466c regulates Vegfa gene transcription in the nucleus by targeting the promoter, and expands on our understanding of the role of miRNAs well beyond their canonical role., Competing Interests: RNatives Inc has filed patent related miR-466c described in this manuscript. The funder [RNatives Oy] provided support in the form of salaries for authors [PL, TAT, MPT] and research materials, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. This does not alter our adherence to PLOS ONE policies on sharing data and materials.
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- 2022
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17. Saving cognitive outcome data in Alzheimer's disease clinical trials during the COVID-19 pandemic: Commentary on the virtual administration of the ADAS-Cog.
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Schwab NA, DesRuisseaux LA, Weinberg MS, and Arnold SE
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Elderly participants in Alzheimer's disease (AD) clinical trials are at high risk of morbidity and mortality with interpersonal exposure to COVID-19, a situation that is likely to continue for the foreseeable future. Yet, in-person neuropsychological assessments remain the mainstay primary outcomes for clinical trials seeking prevention and cure for AD. The Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog) is among the most commonly used cognitive assessment in AD clinical trials, and though currently lacking specific guidelines for virtual administrations, it can be used remotely with appropriate modifications and considerations. Here we propose a novel method of virtual administration of the ADAS-Cog, which considers workarounds for technological and human limitations imposed when the participant is no longer sitting across from the test administrator., Competing Interests: The authors have no conflicts of interest to disclose., (© 2020 The Authors. Alzheimer's & Dementia: Translational Research & Clinical Interventions published by Wiley Periodicals LLC on behalf of Alzheimer's Association.)
- Published
- 2020
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18. Tuberculosis among Newly Arrived Immigrants and Refugees in the United States.
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Liu Y, Phares CR, Posey DL, Maloney SA, Cain KP, Weinberg MS, Schmit KM, Marano N, and Cetron MS
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- Humans, Mass Screening, United States epidemiology, Emigrants and Immigrants, Latent Tuberculosis diagnosis, Latent Tuberculosis epidemiology, Refugees, Tuberculosis
- Abstract
Rationale: U.S. health departments routinely conduct post-arrival evaluation of immigrants and refugees at risk for tuberculosis (TB), but this important intervention has not been thoroughly studied. Objectives: To assess outcomes of the post-arrival evaluation intervention. Methods: We categorized at-risk immigrants and refugees as having had recent completion of treatment for pulmonary TB disease overseas (including in Mexico and Canada); as having suspected TB disease (chest radiograph/clinical symptoms suggestive of TB) but negative culture results overseas; or as having latent TB infection (LTBI) diagnosed overseas. Among 2.1 million U.S.-bound immigrants and refugees screened for TB overseas during 2013-2016, 90,737 were identified as at risk for TB. We analyzed a national data set of these at-risk immigrants and refugees and calculated rates of TB disease for those who completed post-arrival evaluation. Results: Among 4,225 persons with recent completion of treatment for pulmonary TB disease overseas, 3,005 (71.1%) completed post-arrival evaluation within 1 year of arrival; of these, TB disease was diagnosed in 22 (732 cases/100,000 persons), including 4 sputum culture-positive cases (133 cases/100,000 persons), 13 sputum culture-negative cases (433 cases/100,000 persons), and 5 cases with no reported sputum-culture results (166 cases/100,000 persons). Among 55,938 with suspected TB disease but negative culture results overseas, 37,089 (66.3%) completed post-arrival evaluation; of these, TB disease was diagnosed in 597 (1,610 cases/100,000 persons), including 262 sputum culture-positive cases (706 cases/100,000 persons), 281 sputum culture-negative cases (758 cases/100,000 persons), and 54 cases with no reported sputum-culture results (146 cases/100,000 persons). Among 30,574 with LTBI diagnosed overseas, 18,466 (60.4%) completed post-arrival evaluation; of these, TB disease was diagnosed in 48 (260 cases/100,000 persons), including 11 sputum culture-positive cases (60 cases/100,000 persons), 22 sputum culture-negative cases (119 cases/100,000 persons), and 15 cases with no reported sputum-culture results (81 cases/100,000 persons). Of 21,714 persons for whom treatment for LTBI was recommended at post-arrival evaluation, 14,977 (69.0%) initiated treatment and 8,695 (40.0%) completed treatment. Conclusions: Post-arrival evaluation of at-risk immigrants and refugees can be highly effective. To optimize the yield and impact of this intervention, strategies are needed to improve completion rates of post-arrival evaluation and treatment for LTBI.
- Published
- 2020
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19. Broadly active zinc finger protein-guided transcriptional activation of HIV-1.
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Scott TA, O'Meally D, Grepo NA, Soemardy C, Lazar DC, Zheng Y, Weinberg MS, Planelles V, and Morris KV
- Abstract
Human immunodeficiency virus type 1 (HIV-1) causes a persistent viral infection resulting in the demise of immune regulatory cells. Clearance of HIV-1 infection results in integration of proviral DNA into the genome of host cells, which provides a means for evasion and long-term persistence. A therapeutic compound that specifically targets and sustainably activates a latent HIV-1 provirus could be transformative and is the goal for the "shock-and-kill" approach to a functional cure for HIV-1. Substantial progress has been made toward the development of recombinant proteins that target specific genomic loci for gene activation, repression, or inactivation by directed mutations. However, most of these modalities are too large or too complex for efficient therapeutic application. We describe here the development and testing of a novel recombinant zinc finger protein transactivator, ZFP-362-VPR, which specifically and potently enhances proviral HIV-1 transcription both in established latency models and activity across different viral clades. Additionally, ZFP-362-VPR-activated HIV-1 reporter gene expression in a well-established primary human CD4
+ T cell latency model and off-target pathways were determined by transcriptome analyses. This study provides clear proof of concept for the application of a novel, therapeutically relevant, protein transactivator to purge cellular reservoirs of HIV-1., Competing Interests: K.V.M., M.S.W., T.A.S., and D.C.L. have been issued a patent on this technology, W02019079819., (© 2020 The Author(s).)- Published
- 2020
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20. Clinical Trials and Tribulations in the COVID-19 Era.
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Weinberg MS, Patrick RE, Schwab NA, Owoyemi P, May R, McManus AJ, Gerber J, Harper DG, Arnold SE, and Forester B
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- Aged, Alzheimer Disease drug therapy, Alzheimer Disease prevention & control, Betacoronavirus, COVID-19, Guidelines as Topic, Humans, SARS-CoV-2, Clinical Trials as Topic methods, Clinical Trials as Topic standards, Coronavirus Infections prevention & control, Neurocognitive Disorders drug therapy, Neurocognitive Disorders prevention & control, Pandemics prevention & control, Pneumonia, Viral prevention & control
- Abstract
Advances in treating and preventing Alzheimer disease and other neurocognitive disorders of aging arise from rigorous preclinical and clinical research, with randomized controlled treatment trials as the last and definitive test. The COVID-19 pandemic has greatly disrupted ongoing interventional studies and researchers are scrambling to find ways to safely continue this critical work amidst rapidly shifting guidelines from sponsors, institutions, and state and federal guidelines. Here the authors describe novel approaches and work-flow adaptations to study visits, drug delivery and interim and endpoint safety and outcomes assessments to avoid sacrificing years of preparation and substantial financial investments, to work in the best interest of participants and their caregivers, and to continue on the path toward discovering disease-modifying treatments for the millions of individuals impacted by major neurocognitive disorders., (Copyright © 2020 American Association for Geriatric Psychiatry. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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21. An MXD1-derived repressor peptide identifies noncoding mediators of MYC-driven cell proliferation.
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Raffeiner P, Hart JR, García-Caballero D, Bar-Peled L, Weinberg MS, and Vogt PK
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- Aptamers, Nucleotide, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors chemistry, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors genetics, CRISPR-Associated Protein 9 genetics, Capsid Proteins metabolism, Cell Line, Tumor, Clustered Regularly Interspaced Short Palindromic Repeats genetics, Humans, Promoter Regions, Genetic, Protein Domains, RNA, Guide, CRISPR-Cas Systems, RNA, Long Noncoding metabolism, Repressor Proteins chemistry, Repressor Proteins genetics, Transcription, Genetic, Basic Helix-Loop-Helix Leucine Zipper Transcription Factors metabolism, Cell Proliferation genetics, Proto-Oncogene Proteins c-myc metabolism, RNA, Long Noncoding genetics, Repressor Proteins metabolism
- Abstract
MYC controls the transcription of large numbers of long noncoding RNAs (lncRNAs). Since MYC is a ubiquitous oncoprotein, some of these lncRNAs probably play a significant role in cancer. We applied CRISPR interference (CRISPRi) to the identification of MYC-regulated lncRNAs that are required for MYC-driven cell proliferation in the P493-6 and RAMOS human lymphoid cell lines. We identified 320 noncoding loci that play positive roles in cell growth. Transcriptional repression of any one of these lncRNAs reduces the proliferative capacity of the cells. Selected hits were validated by RT-qPCR and in CRISPRi competition assays with individual GFP-expressing sgRNA constructs. We also showed binding of MYC to the promoter of two candidate genes by chromatin immunoprecipitation. In the course of our studies, we discovered that the repressor domain SID (SIN3-interacting domain) derived from the MXD1 protein is highly effective in P493-6 and RAMOS cells in terms of the number of guides depleted in library screening and the extent of the induced transcriptional repression. In the cell lines used, SID is superior to the KRAB repressor domain, which serves routinely as a transcriptional repressor domain in CRISPRi. The SID transcriptional repressor domain is effective as a fusion to the MS2 aptamer binding protein MCP, allowing the construction of a doxycycline-regulatable CRISPRi system that allows controlled repression of targeted genes and will facilitate the functional analysis of growth-promoting lncRNAs., Competing Interests: The authors declare no competing interest., (Copyright © 2020 the Author(s). Published by PNAS.)
- Published
- 2020
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22. Neutralization Breadth and Potency of Single-Chain Variable Fragments Derived from Broadly Neutralizing Antibodies Targeting Multiple Epitopes on the HIV-1 Envelope.
- Author
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van Dorsten RT, Lambson BE, Wibmer CK, Weinberg MS, Moore PL, and Morris L
- Subjects
- Humans, Antibodies, Neutralizing immunology, Antibody Specificity, HIV Antibodies immunology, HIV Envelope Protein gp160 immunology, HIV-1 immunology, Single-Chain Antibodies immunology
- Abstract
Passive administration of HIV-directed broadly neutralizing antibodies (bNAbs) can prevent infection in animal models, and human efficacy trials are under way. Single-chain variable fragments (scFv), comprised of only the variable regions of antibody heavy and light chains, are smaller molecules that may offer advantages over full-length IgG. We designed and expressed scFv of HIV bNAbs prioritized for clinical testing that target the V2-apex (CAP256-VRC26.25), V3-glycan supersite (PGT121), CD4 binding site (3BNC117), and MPER (10E8v4). The use of either a 15- or 18-amino-acid glycine-serine linker between the heavy- and light-chain fragments provided adequate levels of scFv expression. When tested against a 45-multisubtype virus panel, all four scFv retained good neutralizing activity, although there was variable loss of function compared to the parental IgG antibodies. For CAP256-VRC26.25, there was a significant 138-fold loss of potency that was in part related to differential interaction with charged amino acids at positions 169 and 170 in the V2 epitope. Potency was reduced for the 3BNC117 (13-fold) and PGT121 (4-fold) scFv among viruses lacking the N276 and N332 glycans, respectively, and in viruses with a longer V1 loop for PGT121. This suggested that scFv interacted with their epitopes in subtly different ways, with variation at key residues affecting scFv neutralization more than the matched IgGs. Remarkably, the scFv of 10E8v4 maintained breadth of 100% with only a minor reduction in potency. Overall, scFv of clinically relevant bNAbs had significant neutralizing activity, indicating that they are suitable for passive immunization to prevent HIV-1 infection. IMPORTANCE Monoclonal antibodies have been isolated against conserved epitopes on the HIV trimer and are being investigated for passive immunization. Some of the challenges associated with full-sized antibody proteins may be overcome by using single-chain variable fragments (scFv). These smaller forms of antibodies can be produced more efficiently, may show fewer off-target effects with increased tissue penetration, and are more adaptable to vectored-mediated expression than IgG. Here, we demonstrate that scFv of four HIV-directed bNAbs (CAP256-VRC26.25, PGT121, 3BNC117, and 10E8v4) had significant neutralizing activity against diverse global strains of HIV. Loss of potency and/or breadth was shown to be due to increased dependence of the scFv on key residues within the epitope. These smaller antibody molecules with functional activity in the therapeutic range may be suitable for further development as passive immunity for HIV prevention., (Copyright © 2020 American Society for Microbiology.)
- Published
- 2020
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23. AAV-Mediated Expression of Broadly Neutralizing and Vaccine-like Antibodies Targeting the HIV-1 Envelope V2 Region.
- Author
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van den Berg FT, Makoah NA, Ali SA, Scott TA, Mapengo RE, Mutsvunguma LZ, Mkhize NN, Lambson BE, Kgagudi PD, Crowther C, Abdool Karim SS, Balazs AB, Weinberg MS, Ely A, Arbuthnot PB, and Morris L
- Abstract
HIV-1 infection continues to be a global health challenge and a vaccine is urgently needed. Broadly neutralizing antibodies (bNAbs) are considered essential as they inhibit multiple HIV-1 strains, but they are difficult to elicit by conventional immunization. In contrast, non-neutralizing antibodies that correlated with reduced risk of infection in the RV144 HIV vaccine trial are relatively easy to induce, but responses are not durable. To overcome these obstacles, adeno-associated virus (AAV) vectors were used to provide long-term expression of antibodies targeting the V2 region of the HIV-1 envelope protein, including the potent CAP256-VRC26.25 bNAb, as well as non-neutralizing CAP228 antibodies that resemble those elicited by vaccination. AAVs mediated effective antibody expression in cell culture and immunocompetent mice. Mean concentrations of human immunoglobulin G (IgG) in mouse sera increased rapidly following a single AAV injection, reaching 8-60 μg/mL for CAP256 antibodies and 44-220 μg/mL for CAP228 antibodies over 24 weeks, but antibody concentrations varied for individual mice. Secreted antibodies collected from serum retained the expected binding and neutralizing activity. The vectors generated here are, therefore, suitable for the delivery of V2-targeting HIV antibodies, and they could be used in a vectored immunoprophylaxis (VIP) approach to sustain the level of antibody expression required to prevent HIV infection.
- Published
- 2019
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24. Effect of transcription inhibition and generation of suppressive viral non-coding RNAs.
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Pinto DO, Scott TA, DeMarino C, Pleet ML, Vo TT, Saifuddin M, Kovalskyy D, Erickson J, Cowen M, Barclay RA, Zeng C, Weinberg MS, and Kashanchi F
- Subjects
- Anti-Retroviral Agents pharmacology, Biomimetics, CRISPR-Cas Systems, Cell Line, Gene Editing, Gene Silencing, HIV-1 drug effects, Humans, Promoter Regions, Genetic, RNA, Viral genetics, tat Gene Products, Human Immunodeficiency Virus chemistry, Gene Expression Regulation, Viral drug effects, HIV-1 genetics, RNA, Untranslated genetics, Transcription, Genetic
- Abstract
Background: HIV-1 patients receiving combination antiretroviral therapy (cART) survive infection but require life-long adherence at high expense. In chronic cART-treated patients with undetectable viral titers, cell-associated viral RNA is still detectable, pointing to low-level viral transcriptional leakiness. To date, there are no FDA-approved drugs against HIV-1 transcription. We have previously shown that F07#13, a third generation Tat peptide mimetic with competitive activity against Cdk9/T1-Tat binding sites, inhibits HIV-1 transcription in vitro and in vivo., Results: Here, we demonstrate that increasing concentrations of F07#13 (0.01, 0.1, 1 µM) cause a decrease in Tat levels in a dose-dependent manner by inhibiting the Cdk9/T1-Tat complex formation and subsequent ubiquitin-mediated Tat sequestration and degradation. Our data indicate that complexes I and IV contain distinct patterns of ubiquitinated Tat and that transcriptional inhibition induced by F07#13 causes an overall reduction in Tat levels. This reduction may be triggered by F07#13 but ultimately is mediated by TAR-gag viral RNAs that bind suppressive transcription factors (similar to 7SK, NRON, HOTAIR, and Xist lncRNAs) to enhance transcriptional gene silencing and latency. These RNAs complex with PRC2, Sin3A, and Cul4B, resulting in epigenetic modifications. Finally, we observed an F07#13-mediated decrease of viral burden by targeting the R region of the long terminal repeat (HIV-1 promoter region, LTR), promoting both paused polymerases and increased efficiency of CRISPR/Cas9 editing in infected cells. This implies that gene editing may be best performed under a repressed transcriptional state., Conclusions: Collectively, our results indicate that F07#13, which can terminate RNA Polymerase II at distinct sites, can generate scaffold RNAs, which may assemble into specific sets of "RNA Machines" that contribute to gene regulation. It remains to be seen whether these effects can also be seen in various clades that have varying promoter strength, mutant LTRs, and in patient samples.
- Published
- 2019
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25. Acute Pharmacological Management of Behavioral and Emotional Dysregulation Following a Traumatic Brain Injury: A Systematic Review of the Literature.
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Nash RP, Weinberg MS, Laughon SL, McCall RC, Bateman JR, and Rosenstein DL
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- Acute Disease, Brain Injuries, Traumatic psychology, Humans, Practice Guidelines as Topic, Adrenergic beta-Antagonists therapeutic use, Aggression psychology, Amantadine therapeutic use, Anticonvulsants therapeutic use, Brain Injuries, Traumatic drug therapy, Dopamine Agents therapeutic use, Emotional Regulation, Problem Behavior psychology, Propranolol therapeutic use
- Abstract
Background: Traumatic brain injury (TBI) is an increasingly common cause of behavioral and emotional dysregulation among hospitalized patients. While consultation-liaison psychiatrists are often called to help manage these behaviors, acute pharmacological management guidelines are limited., Objective: Conduct a systematic review to determine which pharmacological measures are supported by the literature for targeting agitation and aggression in the acute time period following a TBI., Methods: In a systematic review of MEDLINE, Embase, PsycInfo, ClinicalTrials.gov and the Cochrane Library, we identified and then analyzed publications that investigated the pharmacological management of behavioral and emotional dysregulation following a TBI during the acute time period following injury., Results: There were a limited number of high quality studies that met our inclusion criteria, including only five randomized controlled trials. The majority of the literature identified consisted of case reports or case series. Trends identified in the literature reviewed suggested that amantadine, propranolol, and anti-epileptics were the best supported medications to consider. For many medication classes, the time of medication initiation and duration of treatment, relative to the time of injury, may impact the effect observed., Conclusions: The pharmacological management of agitated patients immediately following a TBI is still an area of much-needed research, as there is limited data-driven guidance in the literature., (Copyright © 2018 Academy of Consultation-Liaison Psychiatry. All rights reserved.)
- Published
- 2019
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26. Stable Transcriptional Repression and Parasitism of HIV-1.
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Shrivastava S, Charlins P, Ackley A, Embree H, Dropulic B, Akkina R, Weinberg MS, and Morris KV
- Abstract
Gene-based therapies represent a promising treatment for HIV-1 infection, as they offer the potential for sustained viral inhibition and reduced treatment interventions. One approach developed here involves using conditionally replicating vectors (CR-vectors). CR-vectors utilize HIV-expressed proteins to replicate and disseminate along with HIV into the budding viral particles, thereby co-infecting target cellular reservoirs. We generated and characterized several CR-vectors carrying various therapeutic payloads of non-coding RNAs targeted to HIV-1, both transcriptionally and post-transcriptionally. Both virus and vector expression was followed in cell culture systems and T cells in the presence and absence of mycophenolic acid (MPA) selection. We find here that CR-vectors functionally suppress HIV expression in a long-term stable manner and that transcriptional targeting of and epigenetic silencing of HIV can be passaged to newly infected cells by the action of the CR-vector, ultimately establishing a sustained parasitism of HIV. Our findings suggest that CR-vectors with modulatory non-coding RNAs may be a viable approach to achieving long-term sustained suppression of HIV-1, leading ultimately to a functional cure., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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27. Letter to the Editor: Integrating Skeletal Muscle Mass and Radiodensity Improves Outcome Prediction and Correlation in Oncologic Studies.
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Williams GR, Weinberg MS, and Shachar SS
- Subjects
- Female, Humans, Muscle, Skeletal, Phenotype, Prognosis, Endometrial Neoplasms, Muscular Diseases
- Published
- 2018
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28. Beyond sarcopenia: Characterization and integration of skeletal muscle quantity and radiodensity in a curable breast cancer population.
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Weinberg MS, Shachar SS, Muss HB, Deal AM, Popuri K, Yu H, Nyrop KA, Alston SM, and Williams GR
- Subjects
- Adult, Aged, Aged, 80 and over, Aging, Body Mass Index, Female, Humans, Middle Aged, Sarcopenia etiology, Tomography, X-Ray Computed, Body Composition physiology, Breast Neoplasms physiopathology, Muscle, Skeletal physiology, Sarcopenia diagnostic imaging
- Abstract
Skeletal muscle loss, commonly known as sarcopenia, is highly prevalent and prognostic of adverse outcomes in oncology. However, there is limited information on adults with early breast cancer and examination of other skeletal muscle indices, despite the potential prognostic importance. This study characterizes and examines age-related changes in body composition of adults with early breast cancer and describes the creation of a novel integrated muscle measure. Female patients diagnosed with stage I-III breast cancer with abdominal computerized tomography (CT) scans within 12 weeks from diagnosis were identified from local tumor registry (N = 241). Skeletal muscle index (muscle area per height [cm
2 /m2 ]), skeletal muscle density, and subcutaneous and visceral adipose tissue areas, were determined from CT L3 lumbar segments. We calculated a novel integrated skeletal measure, skeletal muscle gauge, which combines skeletal muscle index and density (SMI × SMD). 241 patients were identified with available CT imaging. Median age 52 years and range of 23-87. Skeletal muscle index and density significantly decreased with age. Using literature based cut-points, older adults (≥65 years) had significantly higher proportions of sarcopenia (63 vs 28%) and myosteatosis (90 vs 11%) compared to younger adults (<50 years). Body mass index was positively correlated with skeletal muscle index and negatively correlated with muscle density. Skeletal muscle gauge correlated better with increasing age (ρ = 0.52) than with either skeletal muscle index (ρ = 0.20) or density (ρ = 0.46). Wide variations and age-related changes in body composition metrics were found using routinely obtained abdominal CT imaging. Skeletal muscle index and density provide independent, complementary information, and the product of the two metrics, skeletal muscle gauge, requires further research to explore its impact on outcomes in women with curable breast cancer., (© 2017 Wiley Periodicals, Inc.)- Published
- 2018
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29. Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1.
- Author
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Zhou J, Lazar D, Li H, Xia X, Satheesan S, Charlins P, O'Mealy D, Akkina R, Saayman S, Weinberg MS, Rossi JJ, and Morris KV
- Subjects
- Animals, Aptamers, Nucleotide metabolism, Base Sequence, CD4-Positive T-Lymphocytes pathology, CD4-Positive T-Lymphocytes virology, Cell Line, Tumor, DEAD-box RNA Helicases antagonists & inhibitors, DEAD-box RNA Helicases metabolism, Disease Models, Animal, Genetic Therapy methods, HIV Infections immunology, HIV Infections pathology, HIV Infections virology, HIV Long Terminal Repeat, HIV-1 growth & development, HIV-1 metabolism, Humans, Mice, Mice, Inbred NOD, Mice, SCID, Mice, Transgenic, Nucleic Acid Conformation, RNA, Small Interfering genetics, RNA, Small Interfering metabolism, RNA, Viral antagonists & inhibitors, RNA, Viral metabolism, Ribonuclease III antagonists & inhibitors, Ribonuclease III metabolism, Transcription, Genetic, Aptamers, Nucleotide genetics, DEAD-box RNA Helicases genetics, Gene Expression Regulation, Viral, Gene Silencing, HIV Infections therapy, HIV-1 genetics, RNA, Viral genetics, Ribonuclease III genetics
- Abstract
Gene-based therapies represent a promising therapeutic paradigm for the treatment of HIV-1, as they have the potential to maintain sustained viral inhibition with reduced treatment interventions. Such an option may represent a long-term treatment alternative to highly active antiretroviral therapy. Methods: We previously described a therapeutic approach, referred to as transcriptional gene silencing (TGS), whereby small noncoding RNAs directly inhibit the transcriptional activity of HIV-1 by targeting sites within the viral promoter, specifically the 5' long terminal repeat (LTR). TGS differs from traditional RNA interference (RNAi) in that it is characterized by concomitant silent-state epigenetic marks on histones and DNA. To deliver TGS-inducing RNAs, we developed functional RNA conjugates based on the previously reported dual function of the gp120 (A-1) aptamer conjugated to 27-mer Dicer-substrate anti-HIV-1 siRNA (dsiRNA), LTR-362. Results: We demonstrate here that high levels of processed guide RNAs localize to the nucleus in infected T lymphoblastoid CEM cell line and primary human CD4+ T-cells. Treatment of the aptamer-siRNA conjugates induced TGS with an ~10-fold suppression of viral p24 levels as measured at day 12 post infection. To explore the silencing efficacy of aptamer-siRNA conjugates in vivo , HIV-1-infected humanized NOD/SCID/IL2 rγ
null mice (hu-NSG) were treated with the aptamer-siRNA conjugates. Systemic delivery of the A-1-stick-LTR-362 27-mer siRNA conjugates suppressed HIV-1 infection and protected CD4+ T cell levels in viremia hu-NSG mice. Principle conclusions: Collectively these data suggest that the gp120 aptamer-dsiRNA conjugate design is suitable for systemic delivery of small RNAs that can be used to suppress HIV-1., Competing Interests: Conflict of Interest: J. J. R. and J. Z. have an issued patent entitled “Cell-type specific aptamer-siRNA delivery system for HIV-1 therapy”. USPTO, No. US 8, 222, 226 B2, issued date: July 17, 2012. J.J.R., J. Z., M.S.W and K.V.M. have an issued patent entitled “Cell-specific internalizing RNA aptamers against human CCR5 and used therefore”, USPTO, No. US 9,605,266, issued date: March 28, 2017.- Published
- 2018
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30. The impact of skeletal muscle on the pharmacokinetics and toxicity of 5-fluorouracil in colorectal cancer.
- Author
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Williams GR, Deal AM, Shachar SS, Walko CM, Patel JN, O'Neil B, McLeod HL, Weinberg MS, Choi SK, Muss HB, and Sanoff HK
- Subjects
- Adult, Aged, Antibiotics, Antineoplastic administration & dosage, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Bevacizumab administration & dosage, Colorectal Neoplasms diagnostic imaging, Female, Fluorouracil administration & dosage, Fluorouracil therapeutic use, Humans, Leucovorin therapeutic use, Male, Middle Aged, Obesity metabolism, Organoplatinum Compounds therapeutic use, Tomography, X-Ray Computed, Antibiotics, Antineoplastic adverse effects, Antibiotics, Antineoplastic pharmacokinetics, Colorectal Neoplasms drug therapy, Colorectal Neoplasms metabolism, Fluorouracil adverse effects, Fluorouracil pharmacokinetics, Muscle, Skeletal pathology, Sarcopenia complications, Sarcopenia pathology
- Abstract
Purpose: Great heterogeneity exists in the ability of adults with cancer to tolerate chemotherapy. Variability in body composition may affect rates of metabolism of cytotoxic agents and contribute to the variable chemotherapy toxicity observed. The objective of this exploratory study was to examine the association of low skeletal muscle, commonly known as sarcopenia, on the pharmacokinetics (PKs) of 5-fluorouracil (5FU) in patients receiving FOLFOX for colorectal cancer., Methods: We performed a secondary analysis of a completed multicenter trial that investigated PK-guided 5FU dosing in patients receiving mFOLFOX6 +/- bevacizumab for colorectal cancer. Cycle 1 PK samples were obtained 2-44 h after the start of the 5FU infusion (steady state)., Results: No significant differences in first cycle 5FU area-under-the-concentration-time-curve (AUC) were found between sarcopenic and non-sarcopenic patients (17.3 vs. 19.3 AUC, p = 0.43). Patients with grade 3/4 toxicity had a higher dose of 5FU per kg lean body mass (LBM) (105 vs. 93 mg/kg, p = 0.06), most notably for hematological toxicities (110 vs. 94 mg/kg, p = 0.002); however, no correlation between the dose/LBM and 5FU AUC was found., Conclusions: Although our results did not confirm the impact of low skeletal muscle on PKs of 5FU, further research exploring the impact of body composition on chemotherapy PKs and related toxicities is warranted with the potential for alternative dosing strategies in sarcopenic patients to reduce unnecessary toxicities while maintaining efficacy.
- Published
- 2018
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31. Frailty and skeletal muscle in older adults with cancer.
- Author
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Williams GR, Deal AM, Muss HB, Weinberg MS, Sanoff HK, Guerard EJ, Nyrop KA, Pergolotti M, and Shachar SS
- Subjects
- Aged, Cross-Sectional Studies, Female, Frail Elderly statistics & numerical data, Frailty epidemiology, Geriatric Assessment, Humans, Male, Registries, Retrospective Studies, Sarcopenia epidemiology, Frailty diagnosis, Muscle, Skeletal diagnostic imaging, Neoplasms epidemiology, Sarcopenia diagnosis
- Abstract
Objective: Computerized tomography (CT) imaging is routine in oncologic care and can be used to measure muscle quantity and composition that may improve prognostic assessment of older patients. This study examines the association of single-slice CT-assessed muscle measurements with a frailty index in older adults with cancer., Materials and Methods: Using the Carolina Senior Registry, we identified patients with CT imaging within 60days ± of geriatric assessment (GA). A 36-item Carolina Frailty Index was calculated. Cross-sectional skeletal muscle area (SMA) and Skeletal Muscle Density (SMD) were analyzed from CT scan L3 lumbar segments. SMA and patient height (m
2 ) were used to calculate skeletal muscle index (SMI). Skeletal Muscle Gauge (SMG) was calculated by multiplying SMI×SMD., Results: Of the 162 patients, mean age 73, 53% were robust, 27% pre-frail, and 21% frail. Significant differences were found between robust and frail patients for SMD (29.4 vs 24.1 HU, p<0.001) and SMG (1188 vs 922AU, p=0.003), but not SMI (41.9 vs 39.5cm2 /m2 , p=0.29). After controlling for age and gender, for every 5 unit decrease in SMD, the prevalence ratio of frailty increased by 20% (PR=1.20 [1.09, 1.32]) while the prevalence of frailty did not differ based on SMI., Conclusions: Muscle mass (measured as SMI) was poorly associated with a GA-based frailty index. Muscle density, which reflects muscle lipid content, was more associated with frailty. Although frailty and loss of muscle mass are both age-related conditions that are predictive of adverse outcomes, our results suggest they are separate entities., (Copyright © 2017 Elsevier Ltd. All rights reserved.)- Published
- 2018
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32. Molecular trade-offs in RNA ligases affected the modular emergence of complex ribozymes at the origin of life.
- Author
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Dhar N, Weinberg MS, Michod RE, and Durand PM
- Abstract
In the RNA world hypothesis complex, self-replicating ribozymes were essential. For the emergence of an RNA world, less is known about the early processes that accounted for the formation of complex, long catalysts from small passively formed molecules. The functional role of small sequences has not been fully explored and, here, a possible role for smaller ligases is demonstrated. An established RNA polymerase model, the R18, was truncated from the 3' end to generate smaller molecules. All the molecules were investigated for self-ligation functions with a set of oligonucleotide substrates without predesigned base pairing. The smallest molecule that exhibited self-ligation activity was a 40-nucleotide RNA. It also demonstrated the greatest functional flexibility as it was more general in the kinds of substrates it ligated to itself although its catalytic efficiency was the lowest. The largest ribozyme (R18) ligated substrates more selectively and with greatest efficiency. With increase in size and predicted structural stability, self-ligation efficiency improved, while functional flexibility decreased. These findings reveal that molecular size could have increased from the activity of small ligases joining oligonucleotides to their own end. In addition, there is a size-associated molecular-level trade-off that could have impacted the evolution of RNA-based life., Competing Interests: All authors declare no competing interests.
- Published
- 2017
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33. ssAAVs containing cassettes encoding SaCas9 and guides targeting hepatitis B virus inactivate replication of the virus in cultured cells.
- Author
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Scott T, Moyo B, Nicholson S, Maepa MB, Watashi K, Ely A, Weinberg MS, and Arbuthnot P
- Subjects
- CRISPR-Associated Protein 9 metabolism, Gene Editing, Genetic Vectors pharmacology, Hep G2 Cells, Hepatitis B virology, Hepatitis B virus genetics, Hepatitis B virus physiology, Humans, Mutagenesis, Site-Directed, Open Reading Frames, RNA, Guide, CRISPR-Cas Systems chemical synthesis, Staphylococcus aureus metabolism, Virus Replication drug effects, CRISPR-Associated Protein 9 genetics, Dependovirus genetics, Hepatitis B virus drug effects, RNA, Guide, CRISPR-Cas Systems genetics
- Abstract
Management of infection with hepatitis B virus (HBV) remains a global health problem. Persistence of stable covalently closed circular DNA (cccDNA) during HBV replication is responsible for modest curative efficacy of currently licensed drugs. Novel gene editing technologies, such as those based on CRISPR/Cas9, provide the means for permanently disabling cccDNA. However, efficient delivery of antiviral sequences to infected hepatocytes is challenging. A limiting factor is the large size of sequences encoding Cas9 from Streptococcus pyogenes, and resultant incompatibility with the popular single stranded adeno-associated viral vectors (ssAAVs). We thus explored the utility of ssAAVs for delivery of engineered CRISPR/Cas9 of Staphylococcus aureus (Sa), which is encoded by shorter DNA sequences. Short guide RNAs (sgRNAs) were designed with cognates in the S open reading frame of HBV and incorporated into AAVs that also encoded SaCas9. Intended targeted mutation of HBV DNA was observed after transduction of cells with the all-in-one vectors. Efficacy against HBV-infected hNTCP-HepG2 cells indicated that inactivation of cccDNA was successful. Analysis of likely off-target mutagenesis revealed no unintended sequence changes. Use of ssAAVs to deliver all components required to disable cccDNA by SaCas9 is novel and the technology has curative potential for HBV infection.
- Published
- 2017
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34. Skeletal muscle measures and physical function in older adults with cancer: sarcopenia or myopenia?
- Author
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Williams GR, Deal AM, Muss HB, Weinberg MS, Sanoff HK, Nyrop KA, Pergolotti M, and Shachar SS
- Subjects
- Age Factors, Aged, Body Composition, Female, Geriatric Assessment, Humans, Male, Muscle, Skeletal diagnostic imaging, Neoplasms complications, Neoplasms diagnostic imaging, Physical Fitness, Sarcopenia diagnostic imaging, Sarcopenia etiology, Sarcopenia pathology, Sarcopenia physiopathology, Tomography, X-Ray Computed, Muscle, Skeletal pathology, Muscle, Skeletal physiopathology, Neoplasms pathology, Neoplasms physiopathology
- Abstract
Background: Skeletal muscle loss, commonly known as sarcopenia, is highly prevalent in older adults and linked with adverse outcomes in cancer, yet the definition and role of sarcopenia remains uncertain. The aim of this study was to examine the association of Computerized Tomography (CT) assessed skeletal muscle measures with physical function in older adults with cancer., Results: CTs for 185 patients were available. Median age 73 (IQR 68-76) and 56.5% female. After controlling for sex and BMI, we found no evidence that SMI was associated with physical function impairments. Both SMD and SMG were associated physical function impairments and higher values were associated with decreased limitations in instrumental activities of daily living (RR 0.84 [CI 0.73-0.96] and 0.94 [CI 0.89-0.99], respectively), climbing stairs (RR 0.84 [CI 0.76-0.94] and 0.91 [CI 0.87-0.96]), walking 1 block (RR 0.77 [CI 0.67-0.90] and 0.91 [CI 0.85-0.97]), and prolonged Timed Up and Go (RR 0.83 [CI 0.75-0.92] and 0.92 [CI 0.88-0.96])., Materials and Methods: Using the Carolina Senior Registry, we identified patients with CT imaging performed within 60 days +/- of baseline geriatric assessment (GA). Skeletal muscle area and density (SMD) were analyzed from L3 lumbar segments. Muscle area and height (m2) were used to calculate skeletal muscle index (SMI). Skeletal Muscle Gauge (SMG) was created by multiplying SMI x SMD., Conclusions: Skeletal muscle mass as assessed from CT imaging was not associated with physical function impairments. Skeletal muscle radiodensity was more associated with physical function and may aid in identifying older adults at risk for functional impairments.
- Published
- 2017
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35. Viral Vector Reprogramming of Adult Resident Striatal Oligodendrocytes into Functional Neurons.
- Author
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Weinberg MS, Criswell HE, Powell SK, Bhatt AP, and McCown TJ
- Subjects
- Animals, Cell Line, Dependovirus genetics, Humans, Neurons metabolism, Oligodendroglia metabolism, RNA Interference, RNA, Small Interfering, Rats, Cell Transdifferentiation genetics, Cellular Reprogramming genetics, Corpus Striatum cytology, Genetic Vectors genetics, Neurons cytology, Oligodendroglia cytology
- Abstract
Recent advances suggest that in vivo reprogramming of endogenous cell populations provides a viable alternative for neuron replacement. Astrocytes and oligodendrocyte precursor cells can be induced to transdifferentiate into neurons in the CNS, but, in these instances, reprogramming requires either transgenic mice or retroviral-mediated gene expression. We developed a microRNA (miRNA)-GFP construct that in vitro significantly reduced the expression of polypyrimidine tract-binding protein, and, subsequently, we packaged this construct in a novel oligodendrocyte preferring adeno-associated virus vector. Ten days after rat striatal transduction, the vast majority of the GFP-positive cells were oligodendrocytes, but 6 weeks to 6 months later, the majority of GFP-positive cells exhibited neuronal morphology and co-localized with the neuronal marker NeuN. Patch-clamp studies on striatal slices established that the GFP-positive cells exhibited electrophysiological properties indicative of mature neurons, such as spontaneous action potentials and spontaneous inhibitory postsynaptic currents. Also, 3 months after striatal vector administration, GFP-positive terminals in the ipsilateral globus pallidus or substantia nigra retrogradely transported fluorescent beads back to GFP-positive striatal cell bodies, indicating the presence of functional presynaptic terminals. Thus, this viral vector approach provides a potential means to harness resident oligodendrocytes as an endogenous source for in vivo neuronal replacement., (Copyright © 2017 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2017
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36. Trans Women Doing Sex in San Francisco.
- Author
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Williams CJ, Weinberg MS, and Rosenberger JG
- Subjects
- Adult, Aged, Aged, 80 and over, Female, Humans, Male, Middle Aged, San Francisco, Sexual Partners, Young Adult, Sexual Behavior psychology, Sexuality psychology, Transsexualism psychology
- Abstract
This research investigates the sexuality of trans women (individuals who were assigned male status at birth who currently identify as women), by focusing on the "bodily techniques" (Crossley, 2006) they use in "doing" sexuality. The "doing sexuality" framework not only is modeled after the "doing gender" approach of West and Zimmerman (1987), but also utilizes the idea of "sexual embodiment" to emphasize the agency of trans women as they conceptualize and organize their sexuality in a socially recognized way. This is often difficult as they confront discrimination from medical and legal professionals as well as intimate partners who may find it difficult to adapt to the trans woman's atypical body and conception of gender. However, with a study group of 25 trans women from San Francisco, we found the study participants to be adept at overcoming such hurdles and developing techniques to "do" their sexuality. At the same time, we found trans women's agency constrained by the erotic habitus (Green, 2008) of the wider society. The interplay between innovation and cultural tradition provides an opportunity to fashion a more general model of "doing" sexuality.
- Published
- 2016
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37. Erratum to: Trans Women Doing Sex in San Francisco.
- Author
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Williams CJ, Weinberg MS, and Rosenberger JG
- Published
- 2016
- Full Text
- View/download PDF
38. Transcriptional gene silencing in humans.
- Author
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Weinberg MS and Morris KV
- Subjects
- Animals, Cell Line, Epigenesis, Genetic, Humans, Mammals genetics, Models, Biological, RNA, Antisense metabolism, RNA, Long Noncoding genetics, Gene Silencing, Transcription, Genetic
- Abstract
It has been over a decade since the first observation that small non-coding RNAs can functionally modulate epigenetic states in human cells to achieve functional transcriptional gene silencing (TGS). TGS is mechanistically distinct from the RNA interference (RNAi) gene-silencing pathway. TGS can result in long-term stable epigenetic modifications to gene expression that can be passed on to daughter cells during cell division, whereas RNAi does not. Early studies of TGS have been largely overlooked, overshadowed by subsequent discoveries of small RNA-directed post-TGS and RNAi. A reappraisal of early work has been brought about by recent findings in human cells where endogenous long non-coding RNAs function to regulate the epigenome. There are distinct and common overlaps between the proteins involved in small and long non-coding RNA transcriptional regulatory mechanisms, suggesting that the early studies using small non-coding RNAs to modulate transcription were making use of a previously unrecognized endogenous mechanism of RNA-directed gene regulation. Here we review how non-coding RNA plays a role in regulation of transcription and epigenetic gene silencing in human cells by revisiting these earlier studies and the mechanistic insights gained to date. We also provide a list of mammalian genes that have been shown to be transcriptionally regulated by non-coding RNAs. Lastly, we explore how TGS may serve as the basis for development of future therapeutic agents., (© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.)
- Published
- 2016
- Full Text
- View/download PDF
39. Long Non-coding RNA BGas Regulates the Cystic Fibrosis Transmembrane Conductance Regulator.
- Author
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Saayman SM, Ackley A, Burdach J, Clemson M, Gruenert DC, Tachikawa K, Chivukula P, Weinberg MS, and Morris KV
- Subjects
- Cystic Fibrosis metabolism, DNA-Binding Proteins metabolism, Genetic Loci, Humans, Models, Biological, Protein Binding, Cystic Fibrosis genetics, Cystic Fibrosis Transmembrane Conductance Regulator genetics, Gene Expression Regulation, RNA, Antisense genetics, RNA, Long Noncoding
- Abstract
Cystic fibrosis (CF) is a life-shortening genetic disease. The root cause of CF is heritable recessive mutations that affect the cystic fibrosis transmembrance conductance regulator (CFTR) gene and the subsequent expression and activity of encoded ion channels at the cell surface. We show that CFTR is regulated transcriptionally by the actions of a novel long noncoding RNA (lncRNA), designated as BGas, that emanates from intron 11 of the CFTR gene and is expressed in the antisense orientation relative to the protein coding sense strand. We find that BGas functions in concert with several proteins including HMGA1, HMGB1, and WIBG to modulate the local chromatin and DNA architecture of intron 11 of the CFTR gene and thereby affects transcription. Suppression of BGas or its associated proteins results in a gain of both CFTR expression and chloride ion function. The observations described here highlight a previously underappreciated mechanism of transcriptional control and suggest that BGas may serve as a therapeutic target for specifically activating expression of CFTR.
- Published
- 2016
- Full Text
- View/download PDF
40. A viral kinase mimics S6 kinase to enhance cell proliferation.
- Author
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Bhatt AP, Wong JP, Weinberg MS, Host KM, Giffin LC, Buijnink J, van Dijk E, Izumiya Y, Kung HJ, Temple BR, and Damania B
- Subjects
- Computer Simulation, HEK293 Cells, Human Umbilical Vein Endothelial Cells, Humans, Models, Molecular, Ribosomal Protein S6 Kinases, 70-kDa chemistry, Substrate Specificity, Viral Proteins chemistry, Herpesvirus 8, Human enzymology, Ribosomal Protein S6 Kinases, 70-kDa metabolism, Viral Proteins metabolism
- Abstract
Viruses depend upon the host cell for manufacturing components of progeny virions. To mitigate the inextricable dependence on host cell protein synthesis, viruses can modulate protein synthesis through a variety of mechanisms. We report that the viral protein kinase (vPK) encoded by open reading frame 36 (ORF36) of Kaposi's sarcoma-associated herpesvirus (KSHV) enhances protein synthesis by mimicking the function of the cellular protein S6 kinase (S6KB1). Similar to S6KB1, vPK phosphorylates the ribosomal S6 protein and up-regulates global protein synthesis. vPK also augments cellular proliferation and anchorage-independent growth. Furthermore, we report that both vPK and S6KB1 phosphorylate the enzyme 6-phosphofructo-2-kinase/fructose-2, 6-bisphosphatase 2 (PFKFB2) and that both kinases promote endothelial capillary tubule formation.
- Published
- 2016
- Full Text
- View/download PDF
41. Potent and Targeted Activation of Latent HIV-1 Using the CRISPR/dCas9 Activator Complex.
- Author
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Saayman SM, Lazar DC, Scott TA, Hart JR, Takahashi M, Burnett JC, Planelles V, Morris KV, and Weinberg MS
- Subjects
- Bacterial Proteins metabolism, Base Sequence, Binding Sites, CRISPR-Associated Protein 9, Cell Line, Clustered Regularly Interspaced Short Palindromic Repeats, Endonucleases metabolism, Gene Expression Regulation, Viral, HIV Infections metabolism, HIV Infections virology, HIV Long Terminal Repeat genetics, Humans, NF-kappa B metabolism, Nucleotide Motifs, Protein Binding, RNA, Guide, CRISPR-Cas Systems genetics, Transcriptional Activation, CRISPR-Cas Systems, HIV-1 physiology, Multiprotein Complexes metabolism, Virus Activation, Virus Latency
- Abstract
HIV-1 provirus integration results in a persistent latently infected reservoir that is recalcitrant to combined antiretroviral therapy (cART) with lifelong treatment being the only option. The "shock and kill" strategy aims to eradicate latent HIV by reactivating proviral gene expression in the context of cART treatment. Gene-specific transcriptional activation can be achieved using the RNA-guided CRISPR-Cas9 system comprising single guide RNAs (sgRNAs) with a nuclease-deficient Cas9 mutant (dCas9) fused to the VP64 transactivation domain (dCas9-VP64). We engineered this system to target 23 sites within the long terminal repeat promoter of HIV-1 and identified a "hotspot" for activation within the viral enhancer sequence. Activating sgRNAs transcriptionally modulated the latent proviral genome across multiple different in vitro latency cell models including T cells comprising a clonally integrated mCherry-IRES-Tat (LChIT) latency system. We detected consistent and effective activation of latent virus mediated by activator sgRNAs, whereas latency reversal agents produced variable activation responses. Transcriptomic analysis revealed dCas9-VP64/sgRNAs to be highly specific, while the well-characterized chemical activator TNFα induced widespread gene dysregulation. CRISPR-mediated gene activation represents a novel system which provides enhanced efficiency and specificity in a targeted latency reactivation strategy and represents a promising approach to a "functional cure" of HIV/AIDS.
- Published
- 2016
- Full Text
- View/download PDF
42. MINCR is not a MYC-induced lncRNA.
- Author
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Hart JR, Weinberg MS, Morris KV, Roberts TC, Janda KD, Garner AL, and Vogt PK
- Subjects
- Humans, Burkitt Lymphoma metabolism, Gene Expression Regulation, Neoplastic, Gene Regulatory Networks, Lymphoma, B-Cell metabolism, Proto-Oncogene Proteins c-myc metabolism, RNA, Long Noncoding metabolism
- Published
- 2016
- Full Text
- View/download PDF
43. Design of Effective Primary MicroRNA Mimics With Different Basal Stem Conformations.
- Author
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van den Berg FT, Rossi JJ, Arbuthnot P, and Weinberg MS
- Abstract
Primary microRNA (pri-miRNA) mimics are important mediators of effective gene silencing and are well suited for sustained therapeutic applications. Pri-miRNA mimics are processed in the endogenous miRNA biogenesis pathway, where elements of the secondary RNA structure are crucial for efficient miRNA production. Cleavage of the pri-miRNA to a precursor miRNA (pre-miRNA) by Drosha-DGCR8 typically occurs adjacent to a basal stem of ~11 bp. However, a number of pri-miRNA structures are expected to contain slightly shorter or longer basal stems, which may be further disrupted in predicted folding of the expressed pri-miRNA sequence. We investigated the function and processing of natural and exogenous RNA guides from pri-miRNAs with various basal stems (9-13 bp), where a canonical hairpin was predicted to be well or poorly maintained in predicted structures of the expressed sequence. We have shown that RNA guides can be effectively derived from pri-miRNAs with various basal stem conformations, while predicted guide region stability can explain the function of pri-miRNA mimics, in agreement with previously proposed design principles. This study provides insight for the design of effective mimics based on naturally occurring pri-miRNAs and has identified several novel scaffolds suitable for use in gene silencing applications.
- Published
- 2016
- Full Text
- View/download PDF
44. Synthetic SiRNA Delivery: Progress and Prospects.
- Author
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Roberts TC, Ezzat K, El Andaloussi S, and Weinberg MS
- Subjects
- Animals, Drug Carriers chemical synthesis, Hydrophobic and Hydrophilic Interactions, RNA, Small Interfering genetics, Drug Carriers chemistry, RNA, Small Interfering chemistry
- Abstract
Small interfering RNA (siRNA) is a powerful tool for modulating gene expression by RNA interference (RNAi). Duplex RNA oligonucleotides induce cleavage of homologous target transcripts, thereby enabling posttranscriptional silencing of potentially any gene. As such, siRNAs may have utility as novel pharmaceuticals for a wide range of diseases. However, a lack of "drug-likeness," physiological barriers, and potential toxicities have meant that systemic delivery of SiRNAs in vivo remains a major challenge. Here we discuss various strategies that have been employed to solve the problem of SiRNA delivery. These include chemical modification of the SiRNA, direct conjugation to bioactive moieties, and nanoparticle formulations.
- Published
- 2016
- Full Text
- View/download PDF
45. Quantification of nascent transcription by bromouridine immunocapture nuclear run-on RT-qPCR.
- Author
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Roberts TC, Hart JR, Kaikkonen MU, Weinberg MS, Vogt PK, and Morris KV
- Subjects
- Bromouracil analogs & derivatives, Cell Line, High-Throughput Nucleotide Sequencing, Humans, Immunoprecipitation, Uridine analogs & derivatives, Reverse Transcriptase Polymerase Chain Reaction, Transcription, Genetic
- Abstract
Nuclear run-on (NRO) is a method that measures transcriptional activity via the quantification of biochemically labeled nascent RNA molecules derived from nuclear isolates. Widespread use of this technique has been limited because of its technical difficulty relative to steady-state total mRNA analyses. Here we describe a detailed protocol for the quantification of transcriptional activity in human cell cultures. Nuclei are first isolated and NRO transcription is performed in the presence of bromouridine. Labeled nascent transcripts are purified by immunoprecipitation, and transcript levels are determined by reverse-transcription quantitative PCR (RT-qPCR). Data are then analyzed using standard techniques described elsewhere. This method is rapid (the protocol can be completed in 2 d) and cost-effective, exhibits negligible detection of background noise from unlabeled transcripts, requires no radioactive materials and can be performed from as few as 500,000 nuclei. It also takes advantage of the high sensitivity, specificity and dynamic range of RT-qPCR.
- Published
- 2015
- Full Text
- View/download PDF
46. A brave new MYC-amplified world.
- Author
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Weinberg MS, Hart JR, and Vogt PK
- Subjects
- Humans, RNA, Untranslated genetics, Transcriptome genetics, Genes, myc genetics, Proto-Oncogene Proteins c-myc genetics
- Published
- 2015
- Full Text
- View/download PDF
47. The therapeutic application of CRISPR/Cas9 technologies for HIV.
- Author
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Saayman S, Ali SA, Morris KV, and Weinberg MS
- Subjects
- Animals, CRISPR-Associated Proteins genetics, Clustered Regularly Interspaced Short Palindromic Repeats genetics, Genetic Therapy methods, HIV-1 genetics, Humans, CRISPR-Associated Proteins therapeutic use, Genetic Therapy trends, HIV Infections genetics, HIV Infections therapy
- Abstract
Introduction: The use of antiretroviral therapy has led to a significant decrease in morbidity and mortality in HIV-infected individuals. Nevertheless, gene-based therapies represent a promising therapeutic paradigm for HIV-1, as they have the potential for sustained viral inhibition and reduced treatment interventions. One new method amendable to a gene-based therapy is the clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein-9 nuclease (Cas9) gene editing system., Areas Covered: CRISPR/Cas9 can be engineered to successfully modulate an array of disease-causing genetic elements. We discuss the diverse roles that CRISPR/Cas9 may play in targeting HIV and eradicating infection. The Cas9 nuclease coupled with one or more small guide RNAs can target the provirus to mediate excision of the integrated viral genome. Moreover, a modified nuclease-deficient Cas9 fused to transcription activation domains may induce targeted activation of proviral gene expression allowing for the purging of the latent reservoirs. These technologies can also be exploited to target host dependency factors such as the co-receptor CCR5, thus preventing cellular entry of the virus., Expert Opinion: The diversity of the CRISPR/Cas9 technologies offers great promise for targeting different stages of the viral life cycle, and have the capacity for mediating an effective and sustained genetic therapy against HIV.
- Published
- 2015
- Full Text
- View/download PDF
48. Cell-specific RNA aptamer against human CCR5 specifically targets HIV-1 susceptible cells and inhibits HIV-1 infectivity.
- Author
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Zhou J, Satheesan S, Li H, Weinberg MS, Morris KV, Burnett JC, and Rossi JJ
- Subjects
- Aptamers, Nucleotide genetics, Aptamers, Nucleotide metabolism, Base Sequence, CD4-Positive T-Lymphocytes drug effects, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes virology, Cell Culture Techniques, Cells, Cultured, HIV Infections immunology, HIV Infections virology, HIV-1 pathogenicity, Humans, Leukocytes, Mononuclear drug effects, Leukocytes, Mononuclear immunology, Leukocytes, Mononuclear virology, Macrophages immunology, Macrophages virology, RNA Interference, RNA, Small Interfering administration & dosage, RNA, Small Interfering genetics, Receptors, CCR5 metabolism, beta Karyopherins antagonists & inhibitors, beta Karyopherins biosynthesis, beta Karyopherins genetics, Aptamers, Nucleotide administration & dosage, HIV Infections drug therapy, HIV-1 drug effects, Macrophages drug effects, Receptors, CCR5 genetics, SELEX Aptamer Technique methods
- Abstract
The C-C chemokine receptor type 5 (CCR5) is a receptor expressed by T cells and macrophages that serves as a coreceptor for macrophage-tropic HIV-1. Loss of CCR5 is associated with resistance to HIV-1. Here, we combine the live-cell-based SELEX with high-throughput sequencing technology to generate CCR5 RNA aptamers capable of specifically targeting HIV-1 susceptible cells (as small interfering RNA [siRNA] delivery agent) and inhibiting HIV-1 infectivity (as antiviral agent) via block of the CCR5 required for HIV-1 to enter cells. One of the best candidates, G-3, efficiently bound and was internalized into human CCR5-expressing cells. The G-3 specifically neutralized R5 virus infection in primary peripheral blood mononuclear cells, and in vivo generated human CD4(+) T cells with a nanomolar inhibitory concentration 50%. G-3 was also capable of transferring functional siRNAs to CCR5-expressing cells. Collectively, the cell-specific, internalizing, CCR5-targeted aptamers and aptamer-siRNA conjugates offer promise for overcoming some of the current challenges of drug resistance in HIV-1 by providing cell-type- or tissue-specific delivery of various therapeutic moieties., (Copyright © 2015 Elsevier Ltd. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
49. HIV Latency and the noncoding RNA therapeutic landscape.
- Author
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Saayman S, Roberts TC, Morris KV, and Weinberg MS
- Subjects
- Animals, Chronic Disease, HIV Infections genetics, HIV-1 genetics, Humans, Molecular Targeted Therapy methods, RNA Interference physiology, Transcription, Genetic, Genetic Therapy methods, HIV Infections therapy, HIV-1 physiology, RNA, Untranslated genetics, Virus Latency genetics
- Abstract
The Human Immunodeficiency Virus (HIV) belongs to the subfamily of lentiviruses that are characterized by long incubation periods and chronic, persistent infection. The virus integrates into the genome of infected CD4+ cells and, in a subpopulation of cells, adopts a transcriptionally silent state, a process referred to a viral latency. This property makes it exceedingly difficult to therapeutically target the virus and eradicate infection. If left untreated, the inexorable demise of the infected individual's immune system ensues, a causal result of Acquired Immunodeficiency Syndrome (AIDS). Latently infected cells provide a reservoir that maintains viral infection indefinitely. In this chapter we explore the role of noncoding RNAs in HIV infection and in the establishment and maintenance of viral latency. Both short and long noncoding RNAs are endogenous modulators of epigenetic regulation in human cells and play an active role in gene expression. Lastly, we explore therapeutic modalities based on expressed RNAs that are capable of countering infection, transcriptionally regulating the virus, and suppressing or activating the latent state.
- Published
- 2015
- Full Text
- View/download PDF
50. Gene therapy and gene transfer approaches to prevent or treat chronic virus infections.
- Author
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Berkout B, Ertl HC, and Weinberg MS
- Subjects
- Animals, Chronic Disease, Genetic Therapy trends, HIV Infections therapy, HIV-1 genetics, HIV-1 immunology, Hepatitis B therapy, Hepatitis C therapy, Humans, Vaccines, DNA therapeutic use, Viral Vaccines genetics, Virus Diseases genetics, Virus Diseases prevention & control, Gene Transfer Techniques trends, Genetic Therapy methods, Viral Vaccines therapeutic use, Virus Diseases therapy
- Published
- 2015
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