Citations

To assess the role of telomerase activity and telomere length in pancreatic CSCs we used different CSC enrichment methods (CD133, ALDH, sphere formation) in primary patient-derived pancreatic cancer cells. We show that CSCs have higher telomerase activity and longer telomeres than bulk tumor cells. Inhibition of telomerase activity, using genetic knockdown or pharmacological inhibitor (BIBR1532), resulted in CSC marker depletion, abrogation of sphere formation in vitro and reduced tumorigenicity in vivo. Furthermore, we identify a positive feedback loop between stemness factors (NANOG, OCT3/4, SOX2, KLF4) and telomerase, which is essential for the self-renewal of CSCs. Disruption of the balance between telomerase activity and stemness factors eliminates CSCs via induction of DNA damage and apoptosis in primary patient-derived pancreatic cancer samples, opening future perspectives to avoid CSC-driven tumor relapse. In the present study, we demonstrate that telomerase regulation is critical for the “stemness” maintenance in pancreatic CSCs and examine the effects of telomerase inhibition as a potential treatment option of pancreatic cancer. This may significantly promote our understanding of PDAC tumor biology and may result in improved treatment for pancreatic cancer patients.

The cardinal signs of systemic metabolic dysfunction, including insulin resistance, hypertension and a systemic pro‐inflammatory state are risk factors for the development of cardiovascular disease, type 2 diabetes, and neurodegenerative conditions, but there is limited information regarding the impact on recovery after spinal cord injury (SCI). To address this gap, we investigated the impact of systemic insulin resistance generated by consumption of a diet high in fat and sucrose, often referred to as a Western Diet, on neuropathophysiological outcomes in a murine compression model of incomplete SCI. Ten‐week‐old female C57BL6 mice were provided a regular diet (RD) or a diet high in fat and sucrose (HFHS) for 7 weeks prior to experimental compression SCI. Mice consuming a Western Diet showed signs of systemic insulin resistance such as evaluated blood glucose. In addition, signs of insulin resistance within the substance of the spinal cord were also observed, including reductions in the abundance of insulin‐like growth factor 1 receptor (IGF‐1R). Impairments in motor recovery evaluated by BMS score and subscore were present in mice consuming HFHS at 14 and 30 days post injury (dpi). Consumption of a diet rich in fat and sucrose also impaired the mean maximal angle achieved on the inclined plane test and resulted in significant impairments in bladder release. Immunochemical analysis of the spinal cord revealed a number of differences in markers of neural injury between the two groups. First, mice consuming HFHS exhibited significant increases in astrogliosis measured by glial fibrillary acidic protein (GFAP) prior to injury, compared to mice consuming a RD. In fact, the prominent increases in GFAP observed in the intact spinal cord of mice consuming HFHS were equivalent to those observed after SCI in mice consuming a regular chow. In the subacute period, markers of microglial/monocyte activation were elevated in all mice with SCI; however, the increase was substantially greater in the spinal cords of mice consuming HFHS. The spinal cord white matter of mice consuming HFHS also showed reductions in the number of oligodendrocyte progenitor and mature myelinating cells prior to SCI and HFHS‐associated reductions in myelin producing cells persisted at 14 and 30 dpi. In addition, loss of serotonergic axons in the spinal cord after SCI was exacerbated in mice consuming HFHS. Growth associated protein 43 (GAP43), a marker of axonal growth cones was elevated after SCI in the spinal cords of mice consuming a RD or a HFHS diet; however, the magnitude of the increase in growth cones was severely impaired in mice consuming HFHS. These findings suggest that consumption of a Western Diet increases the risk of insulin resistance within the central nervous system and impairs neurobehavioral recovery after SCI. These studies highlight insulin resistance as a new mechanism that limits nervous system repair after SCI and as a potential target for intervention to improve functional outcomes. Support or Funding Information NIH R01 NS052741 and The Minnesota State SCI and TBI Research Grant Program. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

Background Hypertension (HTN) has a strong heritable component and the effectiveness of the treatment for HTN has been shown to be influenced by monogenic guidance. Each of the common HTN pharmacotherapies is ~50% effective in the population and layering blood pressure (BP) medications increases side effects and decreases medication adherence. HTN is a complex disease with primary interactions between the heart, vasculature, and kidney. Known and common genetic variants of each these organs have well‐described mechanisms. Despite the established bell‐curve response to treatment, the complexity of HTN, and functional variants of the major organ systems in HTN: no guidance, to date, utilizes a multi‐gene approach to guide BP therapy. Purpose To assess the association between a weighted multi‐gene algorithm and BP control in patients with HTN. Methods Non‐smokers with a family history of HTN were included in the analysis (n=384; age=61.0±0.9, 11% non‐white). 17 genotypes (ADRB1(2 SNPs), ADRB2 (2SNPs), CYP2D6, WNK (3SNPs), SLC12A3, SCNN1A, alpha‐adducin, renin, angiotensin (3 SNPs), ACE, and the angiotensin receptor) were weighted according to previous effect size in the literature (ΔBP when on the target therapy vs. not) as well as the number of peer‐reviewed papers of quality supporting each functional variant. After weighting, each genotype was entered into an algorithm that assessed pairing (e.g. homozygosity of functional ADRB1 is responsive to β‐blockade, heterozygosity is less responsive, and homozygosity of non‐functional is least responsive) within and between organ systems to predict the pharmacotherapy with greatest impact for each individual patient. Each pharmacotherapy was ranked from 1–4 as most to least likely to respond based on the algorithmic assessment of individual patient's genotypes. Three‐years of data was assessed at six‐month intervals for BP and medication history. Current and past pharmacotherapies were also assessed via chart review. Results There was no difference in initial BP at diagnosis between groups matching the top drug recommendation using the multi‐gene weighted algorithm (n=92) vs. those who did not match (n=292). However, from diagnosis to nadir, patients who matched the primary recommendation had a significantly greater drop in BP when compared to patients who did not (ΔSBP= −39.2±2.4 vs. −32.1±1.3mmHg , ΔDBP= −19.4±1.1 vs. −14.0±1.3mmHg , ΔMAP= −24.4±2.1 vs. −19.0±1.2mmHg, respectively, p<0.05 for SBP and DBP). Further, the difference between diagnosis to current 1yr. average BP was lower in the group that matched the top recommendation (ΔSBP= −33.2±2.3 vs. −27.4±1.2mmHg, ΔDBP=−14.8±1.1 vs. −11.5±1.2, ΔMAP= −21.2±2.3 vs. −15.6±1.8, respectively, p<0.05 for SBP and MAP). Conclusions These data suggest an association between a weighted multi‐gene algorithm on the BP response to pharmacotherapy. This magnitude of difference has previously been shown to be associated with a significant reduction in heart attack and stroke risk. Support or Funding Information Geneticure, Inc. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

We recently demonstrated that high dietary fat (HFD) consumption results in a dramatic loss of myelinating cells and their progenitors in the adult CNS (Yoon et al., 2016). However, the mechanism(s) by which HFD drives dysmyelination are unknown. Since epidemiological studies indicate that metabolic syndrome is a key risk factor for multiple sclerosis (MS), a better understanding of how HFD contributes to oligodendrocyte loss and the capacity for myelin regeneration has the potential to highlight new treatment targets. In mice fed HFD, we found significant decreases in vertical activity by 4 wk and significant changes in ambulatory and stereotypy movements by 12 wk in the Open Field Test. HFD‐fed mice also spent less time in the center of the field as opposed to the perimeter, indicating an anxious phenotype. Mitochondrial isolated from the spinal cord of mice consuming HFD for 4 wk had significantly higher state 3 and state 4 respiration as determined by high‐resolution respirometry. However, by 12 wk, state 4 mitochondrial respiration was significantly less in the HFD group. Targeted LC‐LC‐mass spectroscopy approaches demonstrated that Citric Acid Cycle intermediates are also depleted in the spinal cord of mice consuming HFD for 12 wk compared to controls. Changes in genes related to mitochondrial morphology, quality control processes, and oxidative stress response were also found. In addition, there were significantly fewer Olig2+ cells and increased signs of lipid peroxidation products (4‐Hydroxynonenal‐immunoreactivity) in the corpus callosum of HFD consuming mice compared to those fed a regular diet. Supporting in vivo findings that oligodendrocytes are reduced in the spinal cord of mice consuming HFD, we observed increased mitochondrial fragmentation and a reduced oxygen consumption rate in oligodendrocyte precursors cultured with palmitic acid. Astrocytes however, significantly increased oxygen consumption following exposure to palmitic acid. Together, these findings suggest that changes in the adult spinal cord in response to HFD consumption creates an environment that is less conducive to neural repair by impacting mitochondria function and morphology and impairing oligodendrocyte function. To determine any impact of HFD on myelin regeneration, studies are ongoing to examine myelin regeneration using a murine model of Cuprizone‐mediated demyelination. Neurobehavioral assessments to date reveal significant interactions between HFD and myelin injury and repair. Understanding the cellular environment in response to dietary factors such as HFD consumption can help to guide care for individuals with MS and other conditions in which myelin loss plays a pathological role and has potential to identify new targets for intervention. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .

During the past two decades, anatomy instruction has changed due to curricular reforms in medical schools. Reduction in total hours of anatomy course and cadaver dissection, as well as shifts towards an integrated curricula has raised questions whether changes have had a negative impact on the students’ knowledge of anatomy. Our purpose was to investigate the effect of changes in course hours, curriculum type, and laboratory experience on USMLE Step 1 and Step 2 CK scores. Results of 2009 AAA online survey of anatomy course directors from 54 LCME‐accredited schools were matched with USMLE scores for 6,411 examinees taking Step 1 for the first time in 2009. The USMLE scores included Step 1 total test scores, Step 1 gross anatomy sub‐scores, and Step 2 CK total test scores. Analyses of data indicate that for all three USMLE scores, examinees from schools with stand‐alone gross anatomy courses slightly outperformed examinees from schools with integrated curricula. Conversely, the number of course hours had no effect on how students performed on Step 1 or Step 2 CK. Examinees that were exposed to both dissection and prosection slightly outperformed on Step 2 examinees that had dissection only. While these relationships are statistically significant, they may not be practically meaningful and more research is needed to fully explore different aspects of anatomy curricula and retention of anatomy knowledge.

Objectives Betaine‐homocysteine methyltransferase (BHMT) catalyzes the remethylation of homocysteine. BHMT in the human liver is responsible for up to 25% of homocysteine metabolism. Knowledge of the relationship between genetic polymorphisms and BHMT phenotypes could enhance our understanding of the role of this reaction in drug metabolism. Methods Levels of BHMT enzyme activity and immunoreactive protein were measured in 268 human liver biopsies. Twelve tag BHMT SNPs from our own gene resequencing and HapMap data were used to genotype DNA from these samples. Genotype‐phenotype association analysis was then performed. BHMT enzyme activity averaged 0.077 ± 0.028 units/μg, with a 12 fold variation. Results BHMT protein levels averaged 6.9 ± 3.3 units/μg, with a 50‐fold variation. Three SNPs (rs41272270, rs16876512, and rs6875201), located 28kb upstream, in the 5′‐UTR and in intron 1 of BHMT , respectively, were significantly correlated with both phenotypes (p<10 −8 for enzyme activity and p<10 −5 for protein level). rs41272270 was associated with altered protein binding during electrophoresis mobility shift assay. Conclusions Three tag SNPs were significantly associated with human hepatic BHMT enzyme activity and immunoreactive protein. These observations may help us understand the regulation of BHMT in the human liver and its role in drug metabolism. Supported by NIH U01 GM61388, R01 GM28157, R01 CA132780.

The muscle wasting and impaired muscle function associated with critical llness in ICU patients have negative consequences for recovery from primary disease, but are also the most persistent and debilitating of problems for ICU survivors years after hospital discharge. This study aims at improving our understanding of the mechanisms underlying the muscle wasting in ICU patients by conducting time‐resolved analyses of skeletal muscle structure and function together with measurements of protein synthesis rate, myofibrillar protein and gene expression and intracellular signaling in response to post‐synaptic block of neuromuscular transmission (NMB), sedation, mechanical ventilation and muscle unloading at durations varying from hours to two wks by using a unique experimental rat ICU model. Skeletal muscle structure and function were well maintained during the first four days, followed by a dramatic and progressive decline in muscle size and function. The preferential loss of the molecular motor protein myosin and impaired forcegenerating capacity (specif force) are the major causes underlying the impaired skeletal muscle function in the ICU model. Muscle unloading due to lack of weightbearing (external unloading) and muscle membrane depolarization (internal unloading) are forwarded as very important underlying factors. Supported by the Sweish Research Council and AFM

7552 Background: Axi-cel is approved in the US for the treatment of adult patients with relapsed or refractory LBCL after 2 or more lines of systemic therapy. Post-market long term follow up study of commercial Axi-cel recipients using the Center for International Blood and Marrow Transplant Research was recently completed. Methods: From October 2017 to August 2020, 1,500 Axi-cel recipients from 79 centers were enrolled. Of these, 1001 patients with at least 6 months of follow-up were included in this analysis. Outcomes include complete and overall responses rates (CR and ORR), duration of response (DOR), progression-free and overall survival (PFS and OS), cytokine release syndrome (CRS) (Lee D 2014 and American Society for Transplantation and Cellular Therapy [ASTCT]), immune effector cell associated neurotoxicity syndrome (ICANS), hematologic recovery and subsequent neoplasm (SN). Subgroup analysis by sensitivity to therapy, defined as responsive to the last line of therapy prior to Axi-cel. Median follow-up was 12 months (range, 6-28 months). Results: The median age overall was 62 years, 37% were ≥ 65 years, 83% with Eastern Cooperative Oncology Group (ECOG) performance score 0-1, 28% with transformed lymphoma, 14% with high grade lymphoma, 29% with prior autologous transplant, and 66% with chemotherapy-resistant disease prior to Axi-cel. The median time from diagnosis to Axi-cel infusion was 15 months. Best ORR was 70% (CR 53%). Landmark analysis of patients in CR at 6 months post Axi-cel demonstrates a low number of subsequent progression/death events. With respect to outcomes for chemotherapy-sensitive disease versus resistant disease, the ORR, CR, 12-month PFS and OS were 78% vs. 66%, 60% vs. 48%, 55% (95% CI, 48-62%) vs. 40% (95% CI, 37-44%), and 70% (95% CI, 63-76%) vs. 54% (95% CI, 50-58%), respectively. CRS of any grade was reported in 83% of patients. Incidence of Grades ≥ 3 CRS was 10% according to Lee et al 2014, and 13% according to ASTCT Consensus Grading. Median time to any grade CRS was 4 days (range, 1-28 days), and 93% of CRS cases resolved with a median duration of 7 days (range, 1-121 days). ICANS were reported in 576 (57%) patients, grade >3 was 26%. The median time to onset of ICANS was 7 days (range, 1-82 days), and 86% resolved with a median duration of 9 days (range, 1 to 115 days). Twenty-nine patients (2.9%) reported SN: hematologic (N = 17), solid tumors (N = 12). Conclusions: This is the largest report on Axi-cel in the real-world setting and demonstrates consistent efficacy outcomes and further characterizes safety outcomes. Patients in CR at 6 months have sustained disease control with low number of relapse events. Although patients with therapy-sensitive disease experience better outcomes than patients with therapy-resistant, the overall outcomes on both groups of patients are favorable.