Arhgef11 is a Rho guanine nucleotide exchange factor previously implicated in kidney injury in the Dahl salt‐sensitive rat (SS‐WT). Through exchange of GDP for GTP, Arhgef11 regulates cytoskeletal ...structure, function, and cell‐cell contacts through a variety of stimuli (e.g., G‐protein coupled receptors, growth factors, and shear stress). Genetic studies and reduced Arhgef11 expression in an SS‐Arhgef11SHR‐minimal congenic strain (SHR allele substituted for S allele) significantly decreased proteinuria, fibrosis, and improved renal hemodynamics, without impacting BP compared to the SS‐WT. We hypothesized that actin cytoskeleton regulation through Arhgef11 impacts functional changes in protein reuptake and sodium handling in proximal tubules. Here, we studied the SS‐Arhgef11−/− rat model at 4–12 weeks of age under low and high salt (0.3% or 2% NaCl). On low‐salt, starting at week 6 and continuing through week 12, SS‐Arhgef11−/− animals (n=17) demonstrated significantly reduced proteinuria (week 6, −43.9.0 ± 13.6 mg/24 hours, p=0.0079 and week 12, −102.8 ± 13.6, p<0.0001) compared to SS‐WT animals (n=16). On high‐salt, beginning at week 6, SS‐Arhgef11−/− animals (n=15) demonstrated a significant attenuation of proteinuria from weeks 8 to 12 (week 8, −83.9.0 ± 13.9 mg/24 hours, p<0.0001 and week 12, −118.2 ± 13.9, p<0.0001), along with a substantial reduction in BP (week 12, −42 ± 5 mm Hg, p<0.0001) compared to SS‐WT (n=17). Kidney pathology indices were all significantly improved in SS‐Arhgef11−/− compared to SS‐WT animals (all p<0.05), including glomerular injury/hypertrophy, tubulointerstitial injury/fibrosis, and immune cell infiltration (via flow cytometry). To better understand the molecular mechanisms associated with renal protection from loss of Arhgef11, both RNA sequencing and MS/MS proteomics was performed on kidney from SS‐Arhgef11−/− and SS‐WT at week 4 (before onset of renal injury/proteinuria between groups) and at week 12 (low‐salt). At week 4, 416 genes were differentially expressed between groups, whereas 1017 genes and 363 proteins were observed at week 12 (low‐salt). As anticipated, the ‐omics datasets suggest that loss of Arhgef11 (SS‐Arhgef11−/−) initiates early transcriptome/protein changes in the actin cytoskeleton. Of note, reuptake (endocytosis) of filtered albumin by megalin and cubilin and a large number of solute carrier genes (n=57–63, 2.2 fold‐enrichment (4.22×10−12) were upregulated. Many of the genes and proteins in this pathway are involved in tubular sodium transport (Slc5a= Na+/glucose; Slc6a= Na+/Cl−; Slc8a=Na+/Ca2+; Slc9a3= Na+/H+; and Slc34a= Na+‐Pi). In summary, in vivo phenotyping and multi‐omics techniques provides strong evidence that increased Arhgef11 expression in the Dahl S rat leads to actin cytoskeleton mediated changes in cell morphology and cell function (impaired reuptake filtered protein) that promote kidney injury, hypertension, and decline in kidney function.
Support or Funding Information
1R01HL137673 (MRG). The work performed through the UMMC Molecular and Genomics Facility is supported, in part, by funds from the NIGMS, including P20GM103476, P20GM104357, and P20GM121334.
This is from the Experimental Biology 2019 Meeting. There is no full text article associated with this published in The FASEB Journal.
10043
Background: Our experience of B7H3 chimeric antigen receptor T cells (CAR-T) revealed modest engraftment (max 4.9 cells/uL) and no objective responses on first infusion. Previous CD19 CAR T ...cell trials have revealed that host B Cell CD19 can drive robust engraftment and persistence even in the absence of malignant CD19 expression. We therefore developed a bispecific B7H3xCD19 CAR T cell with the intent of using host CD19 expression to drive better engraftment of the B7H3-targeting CAR. Methods: Young patients with relapsed/refractory solid tumors (R/RST) were enrolled onto a Phase 1 trial (NCT04483778) to examine the safety of autologous T cells lentivirally transduced to express either or both scFV-IgG4hinge-CD28tm-4-1BB-zeta B7H3-specific and CD19-specific CARs. The B7H3 CAR construct contained the methotrexate resistance/selection cassette DHFRdm and the tracking/suicide construct EGFRt and the CD19 CAR construct containing the tracking/suicide construct HER2tg, allowing for differential CAR tracking. CAR T cells were cultured in low-dose methotrexate to select for B7H3 containing CAR T cells and limit exposure to monospecific CD19 CAR T cells. All patients received lymphodepleting fludarabine and cyclophosphamide prior to infusion of cryopreserved CAR-T at the prescribed dose level. The maximal tolerated dose or biologically effective dose (BED) was determined based upon observed toxicity through day 28 from initial CAR-T infusion and using a 3+3 statistical design. Results: 11 subjects (age 5-23, median 18 years) enrolled and received dose level (DL) 0B (0.5 x 10
6
TOTAL CAR-T/kg, n = 7) or DL1B (1 x 10
6
TOTAL CAR-T cells/kg, n = 4). One subject experienced dose limiting toxicities at DL1 of transaminitis, hyponatremia, worsening of a pericardial effusion and hypoxia. Most common toxicities were cytokine release syndrome (CRS) (n = 4, maximum CTCAE grade 2).No occurrence of ICANS was noted. Maximum total circulating CAR-T expansion on first infusion was 796.3 cells/uL and max B7H3 expansion was 374.2 cells/uL with 7/11 continuing with ongoing detection of CAR T cells in the peripheral blood at time of last follow up (21 days to 8 months). CAR engraftment was predominated by the B7H3 CAR single expressing CAR T cells in all subjects. All subjects developed B cell aplasia which is ongoing in 10/11 subjects at time of last check (range of 21 days to 8 months). Best overall response of Stable Disease was observed in 2 of the 11 subjects infused. Conclusions: B7H3xCD19 CAR T cells robustly engraft and expand in patients with R/RST and are safe, but despite ongoing detection of the CAR T cells, no objective responses were observed. It is likely that additional measures will be required to enhance the activity of CAR-T cells to achieve objective responses. STRIvE-02 Arm C will explore the addition of pembrolizumab post bispecific CAR infusion to evaluate the effect of PD1 blockade on anti-tumor activity. Clinical trial information: NCT04483778 .
One of the visions of precision medicine has been to re-define disease taxonomies based on molecular characteristics rather than on phenotypic evidence. However, achieving this goal is highly ...challenging, specifically in neurology. Our contribution is a machine-learning based joint molecular subtyping of Alzheimer's (AD) and Parkinson's Disease (PD), based on the genetic burden of 15 molecular mechanisms comprising 27 proteins (e.g. APOE) that have been described in both diseases. We demonstrate that our joint AD/PD clustering using a combination of sparse autoencoders and sparse non-negative matrix factorization is reproducible and can be associated with significant differences of AD and PD patient subgroups on a clinical, pathophysiological and molecular level. Hence, clusters are disease-associated. To our knowledge this work is the first demonstration of a mechanism based stratification in the field of neurodegenerative diseases. Overall, we thus see this work as an important step towards a molecular mechanism-based taxonomy of neurological disorders, which could help in developing better targeted therapies in the future by going beyond classical phenotype based disease definitions.
Abstract Spaceflight induces an immune response in astronauts. To better characterize this effect, we generated single-cell, multi-ome, cell-free RNA (cfRNA), biochemical, and hematology data for the ...SpaceX Inspiration4 (I4) mission crew. We found that 18 cytokines/chemokines related to inflammation, aging, and muscle homeostasis changed after spaceflight. In I4 single-cell multi-omics data, we identified a “spaceflight signature” of gene expression characterized by enrichment in oxidative phosphorylation, UV response, immune function, and TCF21 pathways. We confirmed the presence of this signature in independent datasets, including the NASA Twins Study, the I4 skin spatial transcriptomics, and 817 NASA GeneLab mouse transcriptomes. Finally, we observed that (1) T cells showed an up-regulation of FOXP3, (2) MHC class I genes exhibited long-term suppression, and (3) infection-related immune pathways were associated with microbiome shifts. In summary, this study reveals conserved and distinct immune disruptions occurring and details a roadmap for potential countermeasures to preserve astronaut health.
Salivary gland cancer comprises a diverse group of histologic types with different biological behavior. Owing to this heterogeneity, the association of margin status and postoperative adjuvant ...radiotherapy has been poorly studied.
To examine the association between surgical margin status and oncologic outcomes and the subsequent outcome of adjuvant radiotherapy in patients with salivary gland carcinomas.
This cohort study analyzed data from institutional records at Memorial Sloan Kettering Cancer Center from 1985 to 2015. Statistical analysis was completed on October 31, 2020. After exclusions, 837 patients with surgically treated salivary gland carcinoma were identified. Surgical margins and histologic characteristics identified from pathology reports were recorded, with margins classified as negative, close, and positive, and individual histologic types classified into 3 risk groups: low, intermediate, and high.
The outcome of adjuvant radiotherapy was determined in patients with close margins with low- and intermediate-risk histologic type and overall pathologic stage I/II disease.
Disease-specific survival (DSS) and local recurrence-free survival (LRFS) outcomes were calculated using the Kaplan-Meier method. Multivariable analysis was performed using the Cox proportional hazards regression model. A planned subgroup analysis of patients with close margins was conducted.
Among the 837 patients identified, 438 were women (52.3%); median age at surgery was 58 years (range, 6-98). A total of 399 tumors (47.7%) originated from major salivary glands, and 438 (52.3%) from minor salivary glands. Margin positivity rates were not different between minor and major salivary gland tumors. Positive surgical margins were identified in 252 patients (30.1%), with nasal cavity/paranasal sinuses and trachea/larynx subsites as the most common sites. Close margins were recorded in 203 patients (24.3%). Adjuvant radiotherapy was administered in 80.5% (103 of 128) of patients with major salivary gland cancer with positive margins, 58.8% (60 of 102) with close margins, and 30.7% (52 of 169) with negative margins and in 70.2% (87 of 124), 36.6% (37 of 101) , and 19.7% (42 of 213) patients with minor salivary gland cancer. With median follow up time of 57 months (range, 1-363 months), patients with positive margins had poorer DSS and LRFS. However, after controlling for overall stage, histologic risk group, and adjuvant radiotherapy, margin status was not a factor associated with poorer DSS or LRFS. In patients with close margins, low-risk and intermediate-risk histologic type, and overall pathologic stage I/II, patients who did not have adjuvant radiotherapy had comparable local control with those who received adjuvant radiotherapy.
The findings of this cohort study suggest that patients with salivary gland cancer who have either close or positive surgical margins are at increased risk for poorer local control and survival. After controlling for tumor stage, histologic risk group, and the use of adjuvant radiotherapy, margin status was not an independent factor associated with poorer outcome. Subgroup analyses showed that care for patients with close margins with low-risk or intermediate-risk histologic type who have stage I/II cancers might be managed safely without adjuvant radiotherapy.
Located at Dome A, the highest point of the Antarctic plateau, the Chinese Kunlun station is considered to be one of the best ground-based photometric sites because of its extremely cold, dry, and ...stable atmosphere. A target can be monitored from there for over 40 days without diurnal interruption during a polar winter. This makes Kunlun station a perfect site to search for short-period transiting exoplanets. Since 2008, an observatory has existed at Kunlun station, and three telescopes are working there. Using these telescopes, the AST3 project has been carried out over the last 6 yr with a search for transiting exoplanets as one of its key programs (CHESPA). In the austral winters of 2016 and 2017, a set of target fields in the southern continuous viewing zone (CVZ) of TESS were monitored by the AST3-II telescope. In this paper, we introduce the CHESPA and present the first data release containing photometry of 26,578 bright stars ( ). The best photometric precision at the optimum magnitude for the survey is around 2 mmag. To demonstrate the data quality, we also present a catalog of 221 variables with a brightness variation greater than 5 mmag from the 2016 data. Among these variables, 179 are newly identified periodic variables not listed in the AAVSO database (https://www.aavso.org/), and 67 are listed in the Candidate Target List. These variables will require careful attention to avoid false-positive signals when searching for transiting exoplanets. Dozens of new transiting exoplanet candidates will be released in a subsequent paper.
Obesity is strongly associated with insulin resistance (IR), along with mitochondrial dysfunction to metabolically active tissues and increased production of reactive O2 species (ROS). Foods rich in ...antioxidants such as wheat germ (WG), protect tissues from damage due to ROS and modulate some negative effects of obesity. This study examined the effects of WG supplementation on markers of IR, mitochondrial substrate metabolism and innate antioxidant markers in two metabolically active tissues (i.e. liver and heart) of C57BL/6 mice fed a high-fat–high-sucrose (HFS) diet. Male C57BL/6 mice, 6-week-old, were randomised into four dietary treatment groups (n 12 mice/group): control (C, 10 % fat kcal), C+10 % WG, HFS (60 % fat kcal) or HFS+10 % WG (HFS+WG). After 12 weeks of treatment, HFS+WG mice had significantly less visceral fat (−16 %, P=0·006) compared with the HFS group. WG significantly reduced serum insulin (P=0·009), the insulinotropic hormone, gastric inhibitory peptide (P=0·0003), and the surrogate measure of IR, homoeostatic model assessment of IR (P=0·006). HFS diet significantly elevated (45 %, P=0·02) cardiac complex 2 mitochondrial VO2, suggesting increased metabolic stress, whereas WG stabilised this effect to the level of control. Consequently, genes which mediate antioxidant defense and mitochondrial biogenesis (superoxide dismutase 2 (Sod2) and PPARγ coactivator 1-α (Pgc1a), respectively) were significantly reduced (P<0·05) in the heart of the HFS group, whereas WG supplementation tended to up-regulate both genes. WG significantly increased hepatic gene expression of Sod2 (P=0·048) but not Pgc1a. Together, these results showed that WG supplementation in HFS diet, reduced IR and improved cardiac mitochondrial metabolic functions.
The recent advances in artificial intelligence, and particularly large language models (LLM) such as chatGPT (openAI, San Francisco, CA, USA), has initiated extensive discussions in the scientific ...community regarding their potential uses and, more importantly, misuses. The real risk here is that the LLM produced a review report that looks properly balanced but has no specific critical content about the manuscript or the described study. Because it summarises the paper and methodology remarkably well, it could easily be mistaken for an actual review report by those that have not fully read the manuscript. ...it is important that all participants within the peer-review process remain vigilant about the use of LLMs.
PDF
