Comprehensive Analysis of Hypermutation in Human Cancer.

We present an extensive assessment of mutation burden through sequencing analysis of >81,000 tumors from pediatric and adult patients, including tumors with hypermutation caused by chemotherapy, carcinogens, or germline alterations. Hypermutation was detected in tumor types not previously associated with high mutation burden. Replication repair deficiency was a major contributing factor. We uncovered new driver mutations in the replication-repair-associated DNA polymerases and a distinct impact of microsatellite instability and replication repair deficiency on the scale of mutation load. Unbiased clustering, based on mutational context, revealed clinically relevant subgroups regardless of the tumors' tissue of origin, highlighting similarities in evolutionary dynamics leading to hypermutation. Mutagens, such as UV light, were implicated in unexpected cancers, including sarcomas and lung tumors. The order of mutational signatures identified previous treatment and germline replication repair deficiency, which improved management of patients and families. These data will inform tumor classification, genetic testing, and clinical trial design.

Expanding the adenovirus toolbox: reporter viruses for studying the dynamics of human adenovirus replication.

To streamline standard virological assays, we developed a suite of nine fluorescent or bioluminescent replication competent human species C5 adenovirus reporter viruses that mimic their parental wild-type counterpart. These reporter viruses provide a rapid and quantitative readout of various aspects of viral infection and replication based on EGFP, mCherry, or NanoLuc measurement. Moreover, they permit real-time non-invasive measures of viral load, replication dynamics, and infection kinetics over the entire course of infection, allowing measurements that were not previously possible. This suite of replication competent reporter viruses increases the ease, speed, and adaptability of standard assays and has the potential to accelerate multiple areas of human adenovirus research.IMPORTANCEIn this work, we developed a versatile toolbox of nine HAdV-C5 reporter viruses and validated their functions in cell culture. These reporter viruses provide a rapid and quantitative readout of various aspects of viral infection and replication based on EGFP, mCherry, or NanoLuc measurement. The utility of these reporter viruses could also be extended for use in 3D cell culture, organoids, live cell imaging, or animal models, and provides a conceptual framework for the development of new reporter viruses representing other clinically relevant HAdV species.

Bacillus subtilis NrnB is expressed during sporulation and acts as a unique 3'-5' exonuclease.

All cells employ a combination of endo- and exoribonucleases to degrade long RNA polymers to fragments 2-5 nucleotides in length. These short RNA fragments are processed to monoribonucleotides by nanoRNases. Genetic depletion of nanoRNases has been shown to increase abundance of short RNAs. This deleteriously affects viability, virulence, and fitness, indicating that short RNAs are a metabolic burden. Previously, we provided evidence that NrnA is the housekeeping nanoRNase for Bacillus subtilis. Herein, we investigate the biological and biochemical functions of the evolutionarily related protein, B. subtilis NrnB (NrnBBs). These experiments show that NrnB is surprisingly different from NrnA. While NrnA acts at the 5' terminus of RNA substrates, NrnB acts at the 3' terminus. Additionally, NrnA is expressed constitutively under standard growth conditions, yet NrnB is selectively expressed during endospore formation. Furthermore, NrnA processes only short RNAs, while NrnB unexpectedly processes both short RNAs and longer RNAs. Indeed, inducible expression of NrnB can even complement the loss of the known global 3'-5' exoribonucleases, indicating that it acts as a general exonuclease. Together, these data demonstrate that NrnB proteins, which are widely found in Firmicutes, Epsilonproteobacteria and Archaea, are fundamentally different than NrnA proteins and may be used for specialized purposes.

Hyperpolarized 13 C metabolic imaging of the human abdomen with spatiotemporal denoising.

PURPOSE: Improving the quality and maintaining the fidelity of large coverage abdominal hyperpolarized (HP) 13 C MRI studies with a patch based global-local higher-order singular value decomposition (GL-HOVSD) spatiotemporal denoising approach. METHODS: Denoising performance was first evaluated using the simulated [1-13 C]pyruvate dynamics at different noise levels to determine optimal kglobal and klocal parameters. The GL-HOSVD spatiotemporal denoising method with the optimized parameters was then applied to two HP [1-13 C]pyruvate EPI abdominal human cohorts (n = 7 healthy volunteers and n = 8 pancreatic cancer patients). RESULTS: The parameterization of kglobal = 0.2 and klocal = 0.9 denoises abdominal HP data while retaining image fidelity when evaluated by RMSE. The kPX (conversion rate of pyruvate-to-metabolite, X = lactate or alanine) difference was shown to be <20% with respect to ground-truth metabolic conversion rates when there is adequate SNR (SNRAUC > 5) for downstream metabolites. In both human cohorts, there was a greater than nine-fold gain in peak [1-13 C]pyruvate, [1-13 C]lactate, and [1-13 C]alanine apparent SNRAUC . The improvement in metabolite SNR enabled a more robust quantification of kPL and kPA . After denoising, we observed a 2.1 ± 0.4 and 4.8 ± 2.5-fold increase in the number of voxels reliably fit across abdominal FOVs for kPL and kPA quantification maps. CONCLUSION: Spatiotemporal denoising greatly improves visualization of low SNR metabolites particularly [1-13 C]alanine and quantification of [1-13 C]pyruvate metabolism in large FOV HP 13 C MRI studies of the human abdomen.

Simulation of interlayer coupling for electroactive covalent organic framework design.

Porous, stacked two-dimensional covalent organic frameworks (2D COFs) bearing semiconducting linkers can support directional charge transfer across adjacent layers of the COF. To better inform the current and possible future design rules for enhancing electron and hole transport in such materials, an understanding of how linker selection and functionalization affects interlayer electronic couplings is essential. We report electronic structure simulation and analysis of electronic couplings across adjacent linker units and to encapsulated species in functionalized electroactive 2D COFs. The detailed dependence of these electronic couplings on interlayer interactions is examined through scans along key interlayer degrees of freedom and through configurational sampling from equilibrium molecular dynamics on semiempirical potential energy surfaces. Beyond affirming the sensitivity of the electronic coupling to interlayer distance and orientation, these studies offer guidance toward linker functionalization strategies for enhancing charge carrier transport in electroactive 2D COFs.

NrnA is a 5'-3' exonuclease that processes short RNA substrates in vivo and in vitro.

Bacterial RNases process RNAs until only short oligomers (2-5 nucleotides) remain, which are then processed by one or more specialized enzymes until only nucleoside monophosphates remain. Oligoribonuclease (Orn) is an essential enzyme that acts in this capacity. However, many bacteria do not encode for Orn and instead encode for NanoRNase A (NrnA). Yet, the catalytic mechanism, cellular roles and physiologically relevant substrates have not been fully resolved for NrnA proteins. We herein utilized a common set of reaction assays to directly compare substrate preferences exhibited by NrnA-like proteins from Bacillus subtilis, Enterococcus faecalis, Streptococcus pyogenes and Mycobacterium tuberculosis. While the M. tuberculosis protein specifically cleaved cyclic di-adenosine monophosphate, the B. subtilis, E. faecalis and S. pyogenes NrnA-like proteins uniformly exhibited striking preference for short RNAs between 2-4 nucleotides in length, all of which were processed from their 5' terminus. Correspondingly, deletion of B. subtilis nrnA led to accumulation of RNAs between 2 and 4 nucleotides in length in cellular extracts. Together, these data suggest that many Firmicutes NrnA-like proteins are likely to resemble B. subtilis NrnA to act as a housekeeping enzyme for processing of RNAs between 2 and 4 nucleotides in length.

Head Impact Exposure in Female Collegiate Soccer by Activity Type.

Soccer, one of the most popular sports in the world, has one of the highest rates of sports-related concussions. Additionally, soccer players are frequently exposed to nonconcussive impacts from intentionally heading the ball, a fundamental component of the sport. There have been many studies on head impact exposure in soccer, but few focus on soccer practices or practice activities. This study aimed to characterize the frequency and magnitude of head impacts in National Collegiate Athletic Association Division I female soccer practice activities using a custom-fit instrumented mouthpiece. Sixteen players were instrumented over the course of 54 practice sessions. Video analysis was performed to verify all mouthpiece-recorded events and classify practice activities. Category groupings of practice activities include technical training, team interaction, set pieces, position-specific, and other. Differences in head impact rates and peak resultant kinematics were observed across activity types and category groupings. Technical training had the highest impact rate compared to other category groupings. Impacts occurring during set piece activities had the highest mean kinematic values. Understanding drill exposure can help inform coaches on training plans aimed to reduce head impact exposure for their athletes.

Real-World Comparison of Bezlotoxumab to Standard of Care Therapy for Prevention of Recurrent Clostridioides difficile Infection in Patients at High Risk for Recurrence.

BACKGROUND: Bezlotoxumab (BEZ) is a monoclonal antibody used to prevent recurrent Clostridioides difficile infection (rCDI). This study investigates BEZ effectiveness in relation to rCDI and patient-specific risk factors in a real-world setting. METHODS: A matched, retrospective cohort study was conducted from 2015 to 2019 to compare BEZ to historical standard of care (SoC) therapy with vancomycin or fidaxomicin. The primary outcome was incidence of 90-day rCDI. Secondary outcomes were incidence of all-cause hospital readmission and all-cause mortality at 90 days, infusion-related reactions, and incidence of heart failure exacerbation. Baseline confounding was addressed using inverse probability of treatment weighting (IPTW). RESULTS: Overall, 107 participants were included (54 BEZ and 53 SoC). Mean number of prior CDI episodes was 2, median number of risk factors for rCDI was 4, and 28% of participants had severe CDI. Incidence of 90-day rCDI was 11% BEZ vs 43% SoC (P = < .001) and 90-day all-cause readmission was 40% BEZ vs 64% SoC (P = .011). In IPTW-adjusted analyses, BEZ was associated with significantly reduced odds of rCDI (odds ratio [OR], 0.14 [95% confidence interval {CI}: .05-.41]) and all-cause readmission (OR, 0.36 [95% CI: .16-.81]). No safety signals were detected with BEZ use. CONCLUSIONS: BEZ is effective for the prevention of rCDI and reduction in all-cause hospital readmission for patients at high risk for recurrence, supporting current guideline recommendations.

High resolution TCSPC imaging of diffuse light with a one-dimensional SPAD array scanning system.

We report a time-correlated single-photon counting (TCSPC) imaging system based on a line-scanning architecture. The system benefits from the high fill-factor, active area, and large dimension of an advanced CMOS single-photon avalanche diode (SPAD) array line-sensor. A two-dimensional image is constructed using a moving mirror to scan the line-sensor field-of-view (FOV) across the target, to enable the efficient acquisition of a two-dimensional 0.26 Mpixel TCSPC image. We demonstrate the capabilities of the system for TCSPC imaging and locating objects obscured in scattering media - specifically to locate a series of discrete point sources of light along an optical fibre submerged in a highly scattering solution. We demonstrate that by selectively imaging using early arriving photons which have undergone less scattering than later arriving photons, our TCSPC imaging system is able to locate the position of discrete point sources of light than a non-time-resolved imaging system.

Head Impact Kinematics and Brain Deformation in Paired Opposing Youth Football Players.

Head impact exposure is often quantified using peak resultant kinematics. While kinematics describes the inertial response of the brain to impact, they do not fully capture the dynamic brain response. Strain, a measure of the tissue-level response of the brain, may be a better predictor of injury. In this study, kinematic and strain metrics were compared to contact characteristics in youth football. Players on 2 opposing teams were instrumented with head impact sensors to record impact kinematics. Video was collected to identify contact scenarios involving opposing instrumented players (ie, paired contact scenarios) and code contact characteristics (eg, player role, impact location). A previously validated, high-resolution brain finite element model, the atlas-based brain model, was used to simulate head impacts and calculate strain metrics. Fifty-two paired contact scenarios (n = 105 impacts) were evaluated. Lighter players tended to have greater biomechanical metrics compared to heavier players. Impacts to the top of the helmet were associated with lower strain metrics. Overall, strain was better correlated with rotational kinematics, suggesting these metrics may be better predictors of the tissue-level brain response than linear kinematics. Understanding the effect of contact characteristics on brain strain will inform future efforts to improve sport safety.

Characterization of Head Impact Exposure in Women's Collegiate Soccer.

Soccer players are regularly exposed to head impacts by intentionally heading the ball. Evidence suggests repetitive subconcussive head impacts may affect the brain, and females may be more vulnerable to brain injury than males. This study aimed to characterize head impact exposure among National Collegiate Athletic Association women's soccer players using a previously validated mouthpiece-based sensor. Sixteen players were instrumented during 72 practices and 24 games. Head impact rate and rate of risk-weighted cumulative exposure were compared across session type and player position. Head kinematics were compared across session type, impact type, player position, impact location, and ball delivery method. Players experienced a mean (95% confidence interval) head impact rate of 0.468 (0.289 to 0.647) head impacts per hour, and exposure rates varied by session type and player position. Headers accounted for 89% of head impacts and were associated with higher linear accelerations and rotational accelerations than nonheader impacts. Headers in which the ball was delivered by a long kick had greater peak kinematics (all P < .001) than headers in which the ball was delivered by any other method. Results provide increased understanding of head impact frequency and magnitude in women's collegiate soccer and may help inform efforts to prevent brain injury.

An examination of a potential organized motor unit firing rate and recruitment scheme of an antagonist muscle during isometric contractions.

The primary purpose of the present study is to determine if an organized control scheme exists for the antagonist muscle during steady isometric torque. A secondary focus is to better understand how firing rates of the antagonist muscle change from a moderate- to higher-contraction intensity. Fourteen subjects performed two submaximal isometric trapezoid muscle actions of the forearm flexors that included a linearly increasing, steady force at both 40% and 70% maximum voluntary contraction, and linearly decreasing segments. Surface electromyographic signals of the biceps and triceps brachii were collected and decomposed into constituent motor unit action potential trains. Motor unit firing rate versus recruitment threshold, motor unit action potential amplitude versus recruitment threshold, and motor unit firing rate versus action potential amplitude relationships of the biceps brachii (agonist) and triceps brachii (antagonist) muscles were analyzed. Moderate- to-strong relationships (|r| ≥ 0.69) were present for the agonist and antagonist muscles for each relationship with no differences between muscles (P = 0.716, 0.428, 0.182). The y-intercepts of the motor unit firing rate versus recruitment threshold relationship of the antagonist did not increase from 40% to 70% maximal voluntary contractions (P = 0.96), unlike for the agonist (P = 0.009). The antagonist muscle exhibits a similar motor unit control scheme to the agonist. Unlike the agonist, however, the firing rates of the antagonist did not increase with increasing intensity. Future research should investigate how antagonist firing rates adapt to resistance training and changes in antagonist firing rates in the absence of peripheral feedback.NEW & NOTEWORTHY This is the first study to explore a potential motor unit control scheme and quantify changes in firing rates with increasing intensity of an antagonist muscle during isometric contractions. We demonstrate that the antagonist muscle possesses an organized motor unit firing rate and recruitment scheme similar to the agonist muscle during isometric forearm flexion, but unlike the agonist muscle, there was no significant increase in firing rates from a moderate- to higher-intensity isometric contraction.

Inhibition of Human Adenovirus Replication by the Importin α/ß1 Nuclear Import Inhibitor Ivermectin.

Human adenoviruses (HAdV) are ubiquitous within the human population and comprise a significant burden of respiratory illnesses worldwide. Pediatric and immunocompromised individuals are at particular risk for developing severe disease; however, no approved antiviral therapies specific to HAdV exist. Ivermectin is an FDA-approved broad-spectrum antiparasitic drug that also exhibits antiviral properties against a diverse range of viruses. Its proposed function is inhibiting the classical protein nuclear import pathway mediated by importin-α (Imp-α) and -ß1 (Imp-ß1). Many viruses, including HAdV, rely on this host pathway for transport of viral proteins across the nuclear envelope. In this study, we show that ivermectin inhibits HAdV-C5 early gene transcription, early and late protein expression, genome replication, and production of infectious viral progeny. Similarly, ivermectin inhibits genome replication of HAdV-B3, a clinically important pathogen responsible for numerous recent outbreaks. Mechanistically, we show that ivermectin disrupts binding of the viral E1A protein to Imp-α without affecting the interaction between Imp-α and Imp-ß1. Our results further extend ivermectin's broad antiviral activity and provide a mechanistic underpinning for its mode of action as an inhibitor of cellular Imp-α/ß1-mediated nuclear import.IMPORTANCE Human adenoviruses (HAdVs) represent a ubiquitous and clinically important pathogen without an effective antiviral treatment. HAdV infections typically cause mild symptoms; however, individuals such as children, those with underlying conditions, and those with compromised immune systems can develop severe disseminated disease. Our results demonstrate that ivermectin, an FDA-approved antiparasitic agent, is effective at inhibiting replication of several HAdV types in vitro This is in agreement with the growing body of literature suggesting ivermectin has broad antiviral activity. This study expands our mechanistic knowledge of ivermectin by showing that ivermectin targets the ability of importin-α (Imp-α) to recognize nuclear localization sequences, without effecting the Imp-α/ß1 interaction. These data also exemplify the applicability of targeting host factors upon which viruses rely as a viable antiviral strategy.

HIV-associated neurocognitive impairment in stable people living with HIV on ART in rural Tanzania.

OBJECTIVES: Few studies have assessed cognitive impairment among healthy people living with HIV (PLWH) who are stable on antiretroviral treatment (ART) in sub-Saharan Africa. METHODS: We conducted a cross-sectional study among a random sample of stable adult PLWH from rural Tanzania on ART for more than 1 year and without immunological failure or pre-existing neurological disease. We evaluated the prevalence and risk factors for neurocognitive impairment (NCI), assessed through neuropsychological tests, functional and depression questionnaires and defined as a mean Z-score ≤ -1 in two or more cognitive domains. RESULTS: Among 243 participants [median age = 44.3 years (interquartile range: 36-52] and 71% female] we found a rate of NCI of 19.3% (95% confidence interval: 14.8-24.8%). Memory and psychomotor domains demonstrated the highest impairment. Independent predictors of NCI were age and self-reported alcohol use. Other classical risk factors were not associated with HIV-associated NCI. CONCLUSION: Despite effective ART roll-out, NCI remained a prevalent condition in this healthy rural Tanzanian population of PLWH on ART. Age and alcohol use were key risk factors.

An examination of motor unit firing rates during steady torque of maximal efforts with either an explosive or slower rate of torque development.

NEW FINDINGS: What is the central question of this study? The aim was to explore agonist and antagonist motor unit firing rates during maximal efforts performed with either an explosive or a slower rate of torque development. What is the main finding and its importance? The antagonist muscle presented a motor unit firing rate relationship similar to the agonist muscle. Additionally, the motor units of both muscles exhibited higher firing rates during explosive maximal contractions than during maximal contractions performed at a slower rate of torque development. These results could prove useful to future research analysing the effects of age, disease, resistance training and/or fatigue-related alterations to motor unit firing rates. ABSTRACT: The primary purpose of the present study was to examine motor unit (MU) firing rates in agonist and antagonist muscles during periods of steady, maximal efforts using explosive and slower rates of torque development. A secondary purpose was to analyse the MU firing rate versus action potential amplitude relationships of the agonist and antagonist muscles during maximal efforts. Thirteen subjects (mean ± SD; age, 21.2 ± 3.6 years; mass 81.1 ± 21.3 kg; and stature, 177.1±9.9 cm) performed two maximal isometric trapezoid muscle actions of the elbow flexors that included either an explosive or a slower, linearly increasing rate (ramp) of torque development. Surface EMG signals of the biceps brachii (BB) and triceps brachii (TB) muscles were collected and decomposed into their constituent MU action potential trains. The MU firing rate versus action potential amplitude relationships of the BB (agonist) and TB (antagonist) muscles were analysed. Moderate to strong relationships (|r| ≥ 0.65) were present for the explosive and ramp contractions in the agonist and antagonist muscles. Firing rates of smaller and larger MUs were higher during the explosive [mean ± SD; agonist = 18.1 ± 6.9 pulses per second (pps), antagonist = 22.0±3.9 pps] than the ramp (agonist = 14.0 ± 5.1 pps, antagonist = 18.3 ± 4.4 pps) contractions for the agonist (P = 0.013) and antagonist muscles (P = 0.007). The antagonist muscle exhibits a similar MU firing rate versus action potential amplitude relationship to the agonist muscle at maximal efforts. Future research should investigate the effects of short-term resistance training on antagonist firing rates and the involvement of peripheral feedback on firing rates during maximal efforts performed at various rates of torque development.

Characterization of On-Field Head Impact Exposure in Youth Soccer.

The objective of this research was to characterize head impacts with a validated mouthpiece sensor in competitive youth female soccer players during a single season with a validated mouthpiece sensor. Participants included 14 youth female soccer athletes across 2 club-level teams at different age levels (team 1, ages 12-13 y; team 2, ages 14-15 y). Head impact and time-synchronized video data were collected for 66 practices and games. Video data were reviewed to characterize the type and frequency of contact experienced by each athlete. A total of 2216 contact scenarios were observed; heading the ball (n = 681, 30.7%) was most common. Other observed contact scenarios included collisions, dives, falls, and unintentional ball contact. Team 1 experienced a higher rate of headers per player per hour of play than team 2, while team 2 experienced a higher rate of collisions and dives. A total of 935 video-verified contact scenarios were concurrent with recorded head kinematics. While headers resulted in a maximum linear acceleration of 56.1g, the less frequent head-to-head collisions (n = 6) resulted in a maximum of 113.5g. The results of this study improve the understanding of head impact exposure in youth female soccer players and inform head impact exposure reduction in youth soccer.

The Effect of Player Contact Characteristics on Head Impact Exposure in Youth Football Games.

To reduce head impact exposure (HIE) in youth football, further understanding of the context in which head impacts occur and the associated biomechanics is needed. The objective of this study was to evaluate the effect of contact characteristics on HIE during player versus player contact scenarios in youth football. Head impact data and time-synchronized video were collected from 4 youth football games over 2 seasons in which opposing teams were instrumented with the Head Impact Telemetry (HIT) System. Coded contact characteristics included the player's role in the contact, player speed and body position, contact height, type, and direction, and head contact surface. Head accelerations were compared among the contact characteristics using mixed-effects models. Among 72 instrumented athletes, 446 contact scenarios (n = 557 impacts) with visible opposing instrumented players were identified. When at least one player had a recorded impact, players who were struck tended to have higher rotational acceleration than players in striking positions. When both players had a recorded impact, lighter players and taller players experienced higher mean head accelerations compared with heavier players and shorter players. Understanding the factors influencing HIE during contact events in football may help inform methods to reduce head injury risk.

Plasma and intracellular pharmacokinetics of tenofovir disoproxil fumarate and emtricitabine in transgender women receiving feminizing hormone therapy.

BACKGROUND: Transwomen have an increased risk of HIV acquisition compared with other adults. Drug-drug interactions between pre-exposure prophylaxis (PrEP) and gender-affirming therapy are cited as a reason for poor PrEP uptake among transwomen. We evaluated plasma tenofovir and emtricitabine pharmacokinetics and their active intracellular anabolites, tenofovir-diphosphate and emtricitabine-triphosphate, in transwomen receiving feminizing hormones. METHODS: We enrolled HIV-negative transwomen (≥19 years) not receiving PrEP. Participants took oral tenofovir disoproxil fumarate/emtricitabine 300/200 mg daily for 14 days. Plasma was collected at 0 h (pre-dose), 0.5, 1, 2, 3, 4, 6, 8 and 12 h on day 14 post-tenofovir disoproxil fumarate/emtricitabine dose. The plasma AUC0-24 was calculated using the trapezoidal rule and compared with historical HIV-negative cisgender adults as geometric mean ratios (GMRs, 90% CI). Secondarily, tenofovir-diphosphate and emtricitabine-triphosphate from PBMCs collected at 0 h and 12 h were reported descriptively as geometric means (90% CI). Clinical trials registration: NCT03270969. RESULTS: Among 15 transwomen (mean age 32 years), geometric mean tenofovir and emtricitabine plasma AUC0-24 were lower compared with controls: tenofovir, 2.10 versus 2.76 mg·h/L, GMR 0.76 (0.65-0.90), P = 0.01; emtricitabine, 9.15 versus 10.64 mg·h/L, GMR 0.86 (0.75-0.98), P = 0.07. Tenofovir-diphosphate and emtricitabine-triphosphate concentrations were higher than previously reported in the literature: 167.1 (146.6-190.5) fmol/106 cells and 15.4 (13.8-17.3) pmol/106 cells, respectively. CONCLUSIONS: We observed lower plasma tenofovir and emtricitabine concentrations in transwomen compared with historical cisgender adults, yet intracellular tenofovir-diphosphate and emtricitabine-triphosphate concentrations were higher than previously reported in PBMCs. Understanding the differences of PrEP pharmacokinetics in plasma and tissue compartments and the resultant impact on efficacy remains important for transwomen.

An Innovative Immunotherapy Vaccine with Combination Checkpoint Blockade as a First Line Treatment for Glioblastoma in the Context of Current Treatments.

Glioblastoma is a devastating disease with a dismal prognosis. While recent advancements in cancer immunotherapy have led to improvements in treating other types of cancer, patients with glioblastoma have not benefited from these new therapies and techniques. Fortunately, neurosurgeons and oncologists at Washington University School of Medicine conducting a cutting edge clinical trial are looking to overcome these persistent challenges in treating glioblastoma through combining a personalized vaccine with new immunotherapy drugs.

Mimicry of Cellular A Kinase-Anchoring Proteins Is a Conserved and Critical Function of E1A across Various Human Adenovirus Species.

The E1A proteins of the various human adenovirus (HAdV) species perform the critical task of converting an infected cell into a setting primed for virus replication. While E1A proteins differ in both sequence and mechanism, the evolutionary pressure on viruses with limited coding capacity ensures that these proteins often have significant overlap in critical functions. HAdV-5 E1A is known to use mimicry to rewire cyclic AMP (cAMP) signaling by decoupling protein kinase A (PKA) from cellular A kinase-anchoring proteins (AKAPs) and utilizing PKA to its own advantage. We show here that E1As from other species of HAdV also possess this viral AKAP (vAKAP) function and examine how they manipulate PKA. E1A from most species of HAdV examined contain a small AKAP-like motif in their N terminus which targets the docking-dimerization domain of PKA as the binding interface for a conserved protein-protein interaction. This motif is also responsible for an E1A-mediated relocalization of PKA regulatory subunits from the cytoplasm into the nucleus, with species-specific E1A proteins having preference for one particular isoform of PKA subunit over another. Importantly, we showed that these newly characterized vAKAPs can integrate into cAMP-responsive transcription as well as contribute to viral genome replication and infectious progeny production for several distinct HAdV species.IMPORTANCE These data enhance the mechanistic knowledge on how HAdV E1A manipulates cellular PKA to benefit infection. The work establishes that mimicry of AKAPs and subversion of PKA-mediated cAMP signaling are conserved features for numerous human adenoviruses. This study also highlights the molecular determinants conferring selective protein-protein interactions between distinct PKA regulatory subunits and the different E1A proteins of these viruses. Additionally, it further emphasizes the utility of using viral proteins like E1A as tools for studying the molecular biology of cellular regulatory pathways.