In-Press Preview
Abstract
BACKGROUND. Identifying a quantitative biomarker of neuropsychiatric dysfunction in people with HIV (PWH) remains a significant challenge in the neuroHIV field. The strongest evidence to date implicates the role of monocytes in central nervous system (CNS) dysfunction in HIV, yet no study has examined monocyte subsets in blood as a correlate and/or predictor of neuropsychiatric function in virally suppressed PWH. METHODS. In two independent cohorts of virologically suppressed women with HIV (vsWWH; n=25 and n=18), whole blood samples were obtained either in conjunction with neuropsychiatric assessments (neuropsychological [NP] test battery, self-report depression and stress-related symptom questionnaires) or one year prior to assessments. Immune cell subsets were assessed by flow cytometry. RESULTS. A higher proportion of intermediate monocytes (CD14+CD16+) was associated with lower global NP function when assessing monocytes concurrently and approximately one year before (predictive) NP testing. The same pattern was seen for executive function (mental flexibility) and processing speed. Conversely, there were no associations with monocyte subsets and depression or stress-related symptoms. Additionally, we found that a higher proportion of classical monocytes was associated with better cognition. CONCLUSION. Although it is widely accepted that lentiviral infection of the CNS targets cells of monocyte-macrophage-microglial lineage, is associated with an increase in intermediate monocytes in the blood and monocyte migration into brain, the percentage of intermediate monocytes in blood of vsWWH has not been associated with neuropsychiatric outcomes. Our findings provide evidence for a new, easily measured blood-based cognitive biomarker in vsWWH. FUNDING. R01-MH113512, R01-MH113512-S, P30-AI094189, R01-MH112391, R01-AI127142, R00-DA044838, U01-AI35004, and P30-MH075673.
Authors
Rebecca T. Veenhuis, Dionna W. Williams, Erin N. Shirk, Celina Monteiro Abreu, Edna A. Ferreira, Jennifer M. Coughlin, Todd T. Brown, Pauline M. Maki, Kathryn Anastos, Joan W. Berman, Janice E. Clements, Leah H. Rubin
Abstract
Effective SARS-CoV-2 vaccines are urgently needed. While most vaccine strategies have focused on systemic immunization, here we compared the protective efficacy of two adjuvanted subunit vaccines with spike protein S1: an intramuscular (IM)- primed /boosted vaccine and an IM-primed/intranasal (IN)-boosted mucosal vaccine, in rhesus macaques. The IM-alum-only vaccine induced robust binding and neutralizing antibody and persistent cellular immunity systemically and mucosally, while IN boosting with nanoparticles including IL-15 and TLR agonists elicited weaker T-cell and antibody responses, but higher dimeric IgA and IFNa. Nevertheless, following SARS-CoV-2 challenge, neither group showed detectable subgenomic RNA in upper or lower respiratory tracts vs naïve controls, indicating full protection against viral replication. Though mucosal and systemic protective mechanisms may differ, results demonstrate both vaccines can protect against respiratory SARS-CoV-2 exposure. The mucosal vaccine was safe after multiple doses and cleared the input virus more efficiently in the nasal cavity, and thus may act as a potent complementary reinforcing boost for conventional systemic vaccines to provide overall better protection.
Authors
Yongjun Sui, Jianping Li, Roushu Zhang, Sunaina Kiran Prabhu, Hanne Andersen Elyard, David Venzon, Anthony Cook, Renita Elizabeth Brown, Elyse Teow, Jason Velasco, Jack Greenhouse, Tammy Putmon-Taylor, Tracey-Ann V. Campbell, Laurent Pessaint, Ian N. Moore, Laurel A. Lagenaur, James D. Talton, Matthew W. Breed, Joshua Kramer, Kevin W. Bock, Mahnaz Minai, Bianca M. Nagata, Mark G. Lewis, Lai-Xi Wang, Jay A. Berzofsky
Abstract
BACKGROUND. Early diagnosis and treatment are key to the long-term survival of lung cancer patients. Although CT has significantly contributed to the early diagnosis of lung cancer, there are still consequences of excessive or delayed treatment. By improving the sensitivity and specificity of circulating tumour cell (CTC) detection, a new solution was proposed for differentiating benign from malignant pulmonary nodules. METHODS. In this study, we used telomerase reverse transcriptase-based CTC detection (TBCD) to distinguish benign from malignant pulmonary nodules <2 cm and compared this method with the pathological diagnosis as the gold standard. FlowSight and FISH were used to confirm the CTCs detected by TBCD. RESULTS. Our results suggest that CTCs based on TBCD can be used as an independent biomarker to distinguish benign from malignant nodules and are significantly superior to serum tumour markers. When the detection threshold was 1, the detection sensitivity and specificity of CTC diagnosis were 0.854 and 0.839, respectively. For pulmonary nodules ≦1 cm and 1-2 cm, the sensitivity and specificity of CTCs were both higher than 77%. Additionally, the diagnostic ability of CTC-assisted CT was compared by CT detection. The results showed that CT combined with CTCs could significantly improve the differentiation ability of benign and malignant nodules in lung nodules <2 cm, and the sensitivity and specificity could reach 0.899 and 0.839, respectively. CONCLUSIONS. In conclusion, TBCD can effectively diagnose pulmonary nodules and be used as an effective auxiliary diagnostic scheme for CT diagnosis. FUNDING. National Key Research and Development Project grants No. 2019YFC1315700 and No. 2017YFC1308702, CAMS Initiative for Innovative Medicine grants No. 2017-I2M-1-005, and National Natural Science Foundation of China grants No. 81472013.
Authors
Wen Zhang, Xinchun Duan, Zhenrong Zhang, Zhenrong Yang, Changyun Zhao, Chunzi Liang, Zhidong Liu, Shujun Cheng, Kaitai Zhang
Abstract
Pleural fibrosis is defined as an excessive deposition of extracellular matrix that results in destruction of the normal pleural tissue architecture and compromised function. Tuberculous pleurisy, asbestos injury and rheumatoid pleurisy are main causes of pleural fibrosis. Pleural mesothelial cells (PMCs) play a key role in pleural fibrosis. However, detailed mechanisms are poorly understood. Serine/arginine-rich protein SRSF6 belongs to a family of highly conserved RNA-binding splicing-factor proteins. Based on its known functions, SRSF6 should be expected to play a role in fibrotic diseases. However, the role of SRSF6 in pleural fibrosis is totally unknown. In this study, SRSF6 protein was found to be increased in cells of tuberculous pleural effusions (TBPE) from patients, and decellularized TBPE, bleomycin and TGFB1 were confirmed to increase SRSF6 levels in PMCs. In vitro, SRSF6 mediated PMC proliferation and synthesis of the main fibrotic protein COL1A2. In vivo, SRSF6 inhibition prevented mouse experimental pleural fibrosis. Lastly, activated-SMAD2/3, increased-SOX4 and depressed-microRNA-506-3p were associated with SRSF6 up-regulation in PMCs. These observations support a model where SRSF6 induces pleural fibrosis through a cluster pathway including SRSF6/WNT5A and SRSF6/SMAD1/5/9 signaling. In conclusion, we propose inhibition of the splicing factor SRSF6 as a strategy for treatment of pleural fibrosis.
Authors
Li-Mei Liang, Liang Xiong, Pei-Pei Cheng, Shuai-Jun Chen, Xiao Feng, Ya-Ya Zhou, Qian Niu, Meng Wang, Qianlan Chen, Lin-Jie Song, Fan Yu, Xin-Liang He, Fei Xiang, Xiaorong Wang, Hong Ye, Wan-Li Ma
Abstract
Despite the availability of multiple HER2-targeted treatments, therapeutic resistance in HER2+ breast cancer remains a clinical challenge. Intratumor heterogeneity for HER2 and resistance-conferring mutations (e.g., PIK3CA) have been investigated in response and resistance to HER2-targeting agents, while the role of divergent cellular phenotypes and tumor epithelial-stromal cell interactions is less well understood. Here, we assessed the effect of intratumor cellular genetic heterogeneity for ERBB2 copy number and PIK3CA mutation on different types of neoadjuvant HER2-targeting therapies and clinical outcome in HER2+ breast cancer. We found that the frequency of cells lacking HER2 was a better predictor of response to HER2-targeted treatment than intratumor heterogeneity. We also compared the efficacy of different therapies in the same tumor using patient-derived xenograft models of heterogeneous HER2+ breast cancer and single cell approaches. Stromal determinants were better predictors of response than tumor epithelial cells, and we identified alveolar epithelial and fibroblastic reticular cells as well as Lyve1+ macrophages as putative drivers of therapeutic resistance. Our results demonstrate that both pre-existing and acquired resistance to HER2-targeting agents involves multiple mechanisms including the tumor microenvironment. Furthermore, our data also suggest that intratumor heterogeneity for HER2 should be incorporated into treatment design.
Authors
Michalina Janiszewska, Shayna Stein, Otto Metzger Filho, Jennifer Eng, Natalie L. Kingston, Nicholas W. Harper, Inga H. Rye, Maša Alečković, Anne Trinh, Katherine C. Murphy, Elisabetta Marangoni, Simona Cristea, Benjamin Oakes, Eric P. Winer, Ian Krop, Hege G. Russnes, Paul T. Spellman, Elmar Bucher, Zhi Hu, Koei Chin, Joe W Gray, Franziska Michor, Kornelia Polyak
Abstract
Emerging coronaviruses from zoonotic reservoirs including severe acute respiratory syndrome coronavirus (SARS-CoV), Middle East respiratory syndrome coronavirus (MERS-CoV), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have been associated with human-to-human transmission and significant morbidity and mortality. Here we study both intradermal (ID) and intramuscular (IM) two-dose delivery regimens of an advanced synthetic DNA vaccine candidate encoding a full-length MERS-CoV Spike (S) protein, which induced potent binding and neutralizing antibodies, as well as cellular immune responses in rhesus macaques. In a MERS-CoV challenge, all immunized rhesus macaques exhibited reduced clinical symptoms, lowered viral lung load, and decreased severity of pathological signs of disease compared to controls. ID vaccination was dose sparing and more effective in this model at protecting animals from disease. The data support the further study of this vaccine for preventing MERS-CoV infection and transmission, including investigation of such vaccines and simplified delivery routes against emerging coronaviruses.
Authors
Ami Patel, Emma L. Reuschel, Ziyang Xu, Faraz I. Zaidi, Kevin Y. Kim, Dana P. Scott, Janess Mendoza, Stephanie Ramos, Regina Stoltz, Friederike Feldmann, Atsushi Okumura, Kimberly Meade-White, Elaine Haddock, Tina Thomas, Rebecca Rosenke, Jamie Lovaglio, Patrick W. Hanley, Greg Saturday, Kar Muthumani, Heinz Feldmann, Laurent M. Humeau, Kate E. Broderick, David B. Weiner
Abstract
BACKGROUND. Serum creatinine concentrations (SCr) are used to determine the presence and severity of acute kidney injury (AKI). SCr is primarily eliminated by glomerular filtration; however, most mechanisms of AKI in critical illness involve kidney proximal tubules, where tubular secretion occurs. Proximal tubular secretory clearance is not currently estimated in the ICU. Our objective was to estimate the kidney clearance of secretory solutes in critically ill adults. METHODS. We collected matched blood and spot urine samples from 170 ICU patients and from a comparison group of 70 adults with normal kidney function. We measured seven endogenously produced secretory solutes using liquid chromatography-tandem mass spectrometry. We computed a composite secretion score incorporating all seven solutes, and evaluated associations with 28-day major adverse kidney events (MAKE28), defined as doubling of SCr, dialysis dependence, or death. RESULTS. The urine/plasma ratio of six of seven secretory solutes were lower in critically ill patients compared with normal individuals after adjustment for SCr. The composite secretion score was moderately correlated with SCr and cystatin C (r = -0.51 and r = -0.53, respectively). Each standard deviation higher composite secretion score was associated with a 25% lower risk of MAKE28 (95% CI 9% - 38% lower) independent of severity of illness, SCr and tubular injury markers. Higher urine to plasma ratios of individual secretory solutes isovalerylglycine and tiglylglycine were associated with MAKE28 after accounting for multiple testing. CONCLUSIONS. Among critically ill adults, tubular secretory clearance is associated with adverse outcomes and measurement could improve assessment of kidney function and dosing of essential ICU medications. TRIAL REGISTRATION. None. FUNDING. PKB was supported by grants from the Digestive and Kidney Diseases K23DK116967, the University of Washington Diabetes Research Center P30DK017047, and an unrestricted gift to the Kidney Research Institute from the Northwest Kidney Centers. EDS was supported by the Vanderbilt O’Brien Kidney Center (NIDDK 5P30 DK114809-03) The funding sources had no role in design and conduct of the study; collection, management, analysis, and interpretation of the data; and preparation, review, or approval of the manuscript.
Authors
Pavan K. Bhatraju, Xin-Ya Chai, Neha A. Sathe, John Ruzinski, Edward D. Siew, Jonathan Himmelfarb, Andrew N. Hoofnagle, Mark M. Wurfel, Bryan R. Kestenbaum
Abstract
Flow-activated Na+ and HCO3- transport in kidney proximal tubules (PTs) underlies relatively constant fractional reabsorption during changes in glomerular filtration rate (GFR), or glomerulotubular balance (GTB). In view of hypothesized connections of epithelial cilia to flow-sensing, we examined flow-activated transport in three polycystic kidney disease-related mouse models based on inducible conditional knockout (KO) of Pkd1, Pkd2, and Kif3a. PTs. were harvested from mice after gene inactivation but prior to cyst formation, and flow-mediated PTs transport were measured. We confirm that higher flow increases both Na+ and HCO3- absorption in control and observe that this flow effect is preserved in PTs of Pkd1-/- and Kif3a-/-mice. However, flow-activation is absent in Pkd2+/- and Pkd2-/- proximal tubules. In heterozygous (Pkd2+/-) mice, a dopamine receptor (DA1) antagonist (SCH23390) restored transport flow sensitivity. When given chronically, this same antagonist reduced renal cyst formation in Pkd2-/- as evidenced by reduced kidney weight, BUN and the cystic index, when compared to untreated mice. In contrast, SCH23390 did not prevent cyst formation in Pkd1-/- mice. These results indicate that Pkd2 is necessary for normal GTB, and that restoration of flow-activated transport by DA1 antagonist can slow renal cyst formation in Pkd2-/- mice.
Authors
Zhaopeng Du, Xin Tian, Ming Ma, Stefan Somlo, Alan M. Weinstein, Tong Wang
Abstract
Altered mitochondria activity in osteoblasts and osteoclast has been implicated in the loss of bone mass associated with aging and estrogen deficiency – the two most common causes of osteoporosis. However, the mechanisms that control mitochondrial metabolism in bone cells during health or disease remain unknown. The mitochondrial deacetylase Sirtuin-3 (Sirt3) has been earlier implicated in age-related diseases. Here, we show that deletion of Sirt3 had no effect on the skeleton of young mice but attenuated the age-related loss of bone mass in both sexes. This effect was associated with impaired bone resorption. Osteoclast progenitors from aged Sirt3 null mice were able to differentiate into osteoclasts. Albeit, the differentiated cells exhibited impaired polykaryon formation and resorptive activity as well as decreased oxidative phosphorylation and mitophagy. The Sirt3 inhibitor LC-0296 recapitulated the effects of Sirt3 deletion in osteoclast formation and mitochondrial function, and its administration to aging mice increased bone mass. Deletion of Sirt3 also attenuated the increase in bone resorption and loss of bone mass caused by estrogen deficiency. These findings suggest that Sirt3 inhibition and the resulting impairment of osteoclast mitochondrial function could be a novel therapeutic intervention for the two most important causes of osteoporosis.
Authors
Wen Ling, Kimberly Krager, Kimberly K. Richardson, Aaron D. Warren, Filipa Ponte, Nukhet Aykin-Burns, Stavros C. Manolagas, Maria Almeida, Ha-Neui Kim
Abstract
Human induced pluripotent stem cell derived cardiomyocytes (hiPSC-CMs) have been used extensively to model inherited heart diseases, but hiPSC-CM models of ischemic heart disease are lacking. Here our objective was to generate an hiPSC-CM model of ischemic heart disease. To this end, hiPSCs were differentiated to functional hiPSC-CMs and then purified using either a simulated ischemia media or by using magnetic antibody-based purification targeting the non-myocyte population for depletion from the cell population. Flow cytometry analysis confirmed that each purification approach generated hiPSC-CM cultures of >94% cTnT+ cells. Following purification hiPSC-CMs were re-plated as confluent syncytial monolayers for electrophysiological phenotype analysis and protein expression by Western blotting. Metabolic selected hiPSC-CM monolayers’ phenotype recapitulated many of the functional and structural hallmarks of ischemic cardiomyocytes, including: elevated diastolic calcium, diminished calcium transient amplitude, prolonged action potential duration, depolarized resting membrane potential, hypersensitivity to chemotherapy induced cardiotoxicity, depolarized mitochondrial membrane potential, depressed SERCA2a expression, reduced maximal oxygen consumption rate and abnormal response to β1-adrenergic receptor stimulation. These findings indicate that metabolic selection of hiPSC-CMs generates cell populations with phenotype like what is well known to occur in the setting of ischemic heart failure, and thus provides a novel opportunity for study of human ischemic heart disease.
Authors
Justin Davis, Ahmad Chouman, Jeffery Creech, Andre Monteiro da Rocha, Daniela Ponce-Balbuena, Eric N. Jimenez Vazquez, Ruthann Nichols, Andrey Lozhkin, Nageswara R. Madamanchi, Katherine F. Campbell, Todd J. Herron
No posts were found with this tag.
