Unresolved Issues in RNA Therapeutics in Vascular Diseases With a Focus on Aneurysm Disease

New technologies have greatly shaped the scientific and medical landscape within the last years. The unprecedented expansion of data and information on RNA biology has led to the discovery of new RNA classes with unique functions and unexpected modifications. Today, the biggest challenge is to transfer the large number of findings in basic RNA biology into corresponding clinical RNA-based therapeutics. Lately, this research begins to yield positive outcomes. RNA drugs advance to the final phases of clinical trials or even receive FDA approval. Furthermore, the introduction of the RNA-guided gene-editing technology CRISPR and advances in the delivery ofmessenger RNAs have triggered a major progression in the field of RNA-therapeutics. Especially short interfering RNAs and antisense oligonucleotides are promising examples for novel categories of therapeutics. However, several issues need to be addressed including intracellular delivery, toxicity, and immune responses before utilizing RNAs in a clinical setting. In this review, we provide an overview on opportunities and challenges for clinical translation of RNA-based therapeutics, with an emphasis on advances in novel delivery technologies and abdominal aortic aneurysm disease where non-coding RNAs have been shown to play a crucial regulatory role

Compartmentalization of androgen receptor protein–protein interactions in living cells

Steroid receptors regulate gene expression in a ligand-dependent manner by binding specific DNA sequences. Ligand binding also changes the conformation of the ligand binding domain (LBD), allowing interaction with coregulators via LxxLL motifs. Androgen receptors (ARs) preferentially interact with coregulators containing LxxLL-related FxxLF motifs. The AR is regulated at an extra level by interaction of an FQNLF motif in the N-terminal domain with the C-terminal LBD (N/C interaction). Although it is generally recognized that AR coregulator and N/C interactions are essential for transcription regulation, their spatiotemporal organization is largely unknown. We performed simultaneous fluorescence resonance energy transfer and fluorescence redistribution after photobleaching measurements in living cells expressing ARs double tagged with yellow and cyan fluorescent proteins. We provide evidence that AR N/C interactions occur predominantly when ARs are mobile, possibly to prevent unfavorable or untimely cofactor interactions. N/C interactions are largely lost when AR transiently binds to DNA, predominantly in foci partly overlapping transcription sites. AR coregulator interactions occur preferentially when ARs are bound to DNA

Variation in Structure and Process of Care in Traumatic Brain Injury: Provider Profiles of European Neurotrauma Centers Participating in the CENTER-TBI Study.

INTRODUCTION: The strength of evidence underpinning care and treatment recommendations in traumatic brain injury (TBI) is low. Comparative effectiveness research (CER) has been proposed as a framework to provide evidence for optimal care for TBI patients. The first step in CER is to map the existing variation. The aim of current study is to quantify variation in general structural and process characteristics among centers participating in the Collaborative European NeuroTrauma Effectiveness Research in Traumatic Brain Injury (CENTER-TBI) study. METHODS: We designed a set of 11 provider profiling questionnaires with 321 questions about various aspects of TBI care, chosen based on literature and expert opinion. After pilot testing, questionnaires were disseminated to 71 centers from 20 countries participating in the CENTER-TBI study. Reliability of questionnaires was estimated by calculating a concordance rate among 5% duplicate questions. RESULTS: All 71 centers completed the questionnaires. Median concordance rate among duplicate questions was 0.85. The majority of centers were academic hospitals (n = 65, 92%), designated as a level I trauma center (n = 48, 68%) and situated in an urban location (n = 70, 99%). The availability of facilities for neuro-trauma care varied across centers; e.g. 40 (57%) had a dedicated neuro-intensive care unit (ICU), 36 (51%) had an in-hospital rehabilitation unit and the organization of the ICU was closed in 64% (n = 45) of the centers. In addition, we found wide variation in processes of care, such as the ICU admission policy and intracranial pressure monitoring policy among centers. CONCLUSION: Even among high-volume, specialized neurotrauma centers there is substantial variation in structures and processes of TBI care. This variation provides an opportunity to study effectiveness of specific aspects of TBI care and to identify best practices with CER approaches

Variation in general supportive and preventive intensive care management of traumatic brain injury: a survey in 66 neurotrauma centers participating in the Collaborative European NeuroTrauma Effectiveness Research in Traumatic Brain Injury (CENTER-TBI) study

Abstract Background General supportive and preventive measures in the intensive care management of traumatic brain injury (TBI) aim to prevent or limit secondary brain injury and optimize recovery. The aim of this survey was to assess and quantify variation in perceptions on intensive care unit (ICU) management of patients with TBI in European neurotrauma centers. Methods We performed a survey as part of the Collaborative European NeuroTrauma Effectiveness Research in Traumatic Brain Injury (CENTER-TBI) study. We analyzed 23 questions focused on: 1) circulatory and respiratory management; 2) fever control; 3) use of corticosteroids; 4) nutrition and glucose management; and 5) seizure prophylaxis and treatment. Results The survey was completed predominantly by intensivists (n = 33, 50%) and neurosurgeons (n = 23, 35%) from 66 centers (97% response rate). The most common cerebral perfusion pressure (CPP) target was > 60 mmHg (n = 39, 60%) and/or an individualized target (n = 25, 38%). To support CPP, crystalloid fluid loading (n = 60, 91%) was generally preferred over albumin (n = 15, 23%), and vasopressors (n = 63, 96%) over inotropes (n = 29, 44%). The most commonly reported target of partial pressure of carbon dioxide in arterial blood (PaCO2) was 36–40 mmHg (4.8–5.3 kPa) in case of controlled intracranial pressure (ICP) < 20 mmHg (n = 45, 69%) and PaCO2 target of 30–35 mmHg (4–4.7 kPa) in case of raised ICP (n = 40, 62%). Almost all respondents indicated to generally treat fever (n = 65, 98%) with paracetamol (n = 61, 92%) and/or external cooling (n = 49, 74%). Conventional glucose management (n = 43, 66%) was preferred over tight glycemic control (n = 18, 28%). More than half of the respondents indicated to aim for full caloric replacement within 7 days (n = 43, 66%) using enteral nutrition (n = 60, 92%). Indications for and duration of seizure prophylaxis varied, and levetiracetam was mostly reported as the agent of choice for both seizure prophylaxis (n = 32, 49%) and treatment (n = 40, 61%). Conclusions Practice preferences vary substantially regarding general supportive and preventive measures in TBI patients at ICUs of European neurotrauma centers. These results provide an opportunity for future comparative effectiveness research, since a more evidence-based uniformity in good practices in general ICU management could have a major impact on TBI outcome

Variation in neurosurgical management of traumatic brain injury

Background: Neurosurgical management of traumatic brain injury (TBI) is challenging, with only low-quality evidence. We aimed to explore differences in neurosurgical strategies for TBI across Europe. Methods: A survey was sent to 68 centers participating in the Collaborative European Neurotrauma Effectiveness Research in Traumatic Brain Injury (CENTER-TBI) study. The questionnaire contained 21 questions, including the decision when to operate (or not) on traumatic acute subdural hematoma (ASDH) and intracerebral hematoma (ICH), and when to perform a decompressive craniectomy (DC) in raised intracranial pressure (ICP). Results: The survey was completed by 68 centers (100%). On average, 10 neurosurgeons work in each trauma center. In all centers, a neurosurgeon was available within 30 min. Forty percent of responders reported a thickness or volume threshold for evacuation of an ASDH. Most responders (78%) decide on a primary DC in evacuating an ASDH during the operation, when swelling is present. For ICH, 3% would perform an evacuation directly to prevent secondary deterioration and 66% only in case of clinical deterioration. Most respondents (91%) reported to consider a DC for refractory high ICP. The reported cut-off ICP for DC in refractory high ICP, however, differed: 60% uses 25 mmHg, 18% 30 mmHg, and 17% 20 mmHg. Treatment strategies varied substantially between regions, specifically for the threshold for ASDH surgery and DC for refractory raised ICP. Also within center variation was present: 31% reported variation within the hospital for inserting an ICP monitor and 43% for evacuating mass lesions. Conclusion: Despite a homogeneous organization, considerable practice variation exists of neurosurgical strategies for TBI in Europe. These results provide an incentive for comparative effectiveness research to determine elements of effective neurosurgical care

MicroRNA-146a-based mechanisms of vascular fibrosis in arterial stiffening

DZHK e.V. (Promotionsstipendium WS16/17)Die altersbedingte arterielle Gefäßversteifung stellt sowohl ein schwerwiegendes Krankheitsbild des Herz-Kreislauf-Systems sowie einen eigenen Risikofaktor für die Entstehung von kardiovaskulären Erkrankungen dar. Die zugrunde liegenden pathologischen Mechanismen sind Gegenstand aktueller Forschung. Besonders Veränderungen in der Extrazellularmatrix der Aorta durch gesteigerte Kollagensynthese oder vermehrte Elastinfragmentation spielen dabei eine Rolle. Es gibt Hinweise, dass ein erhöhtes TGF-β1/SMAD3-Signaling vaskuläre Fibrose auslösen kann. Weiterhin konnten Kollagen- Crosslinks vom HP-Typ vermehrt in fibrotischem Gewebe nachgewiesen werden. MiRs sind post-transkriptionelle Regulatoren, die ganze Gennetzwerke regulieren können. MiR-146a ist bereits als protektiver Regulator von inflammatorischen und fibrogenen Prozessen beschrieben worden. Weiterhin konnte miR-146a bei Patient:innen mit erhöhter arterieller Gefäßversteifung vermehrt im Blut gemessen werden. SMAD3 ist bereits als Zielgen von miR-146a beschrieben worden. Ebenfalls konnte bereits gezeigt werden, dass SMAD3 als Transkriptionsfaktor für das Gen PLOD2 fungiert, welches für die Entstehung HP entscheidend ist. Ziel der vorliegenden Arbeit war es, den Einfluss von miR-146a auf den Mechanismus der gesteigerten Kollagensynthese im Rahmen der altersbedingten arteriellen Gefäßversteifung zu untersuchen. Die Ergebnisse zeigten, dass miR-146a sowohl in alten, murinen Aorten, als auch auf steifen Zellkulturplatten signifikant hochreguliert ist. Die Gene SMAD3 und PLOD2 waren nach Transfektion von miR-146a-Mimic signifikant herunterreguliert und nach Transfektion von miR-146a-Inhibitor signifikant hochreguliert. Die Ergebnisse des Experiments mit siRNA für SMAD3 waren hingegen nicht signifikant. Das Scar-in-a-jar Assay zeigte, dass der miR-146a-Mimic die Synthese von Kollagen vermindert und der miR-146a-Inhibitor die Synthese erhöht. Die vorliegenden Ergebnisse sprechen dafür, dass miR-146a protektiv auf die Entstehung von vaskulärer Fibrose wirkt. Die Regulation könnte über den TGF-β1/SMAD3-Signalweg und über die vermehrte Bildung von Crosslinks durch die LH2 und deren Gen PLOD2 ablaufen. Die erhobenen Daten zeigen erstmalig miR-146a als wichtigen Regulator der vaskulären Fibrose im Rahmen der altersbedingten arteriellen Gefäßversteifung. MiR-146a ist damit vielversprechender Ansatzpunkt für neuartige miR-basierte Therapieansätze der arteriellen Gefäßversteifung.Age-related arterial stiffening is both a serious cardiovascular disease and a risk factor in its own for other cardiovascular diseases. The underlying pathological mechanisms are the subject of current research. In particular, changes in the extracellular matrix of the aorta due to increased collagen synthesis or increased elastin fragmentation seem to be crucial. There is evidence that increased TGF-β1/SMAD3 signaling can trigger vascular fibrosis. Furthermore, collagen crosslinks of the HP type are incresased in fibrotic tissue. MiRs are post-transcriptional regulators that can regulate entire gene networks. MiR-146a has already been described as a protective regulator of inflammatory and fibrogenic processes. Furthermore, miR-146a could be measured in the blood of patients with increased arterial stiffness. SMAD3 has already been described as a target gene of miR-146a. It has also already been shown that SMAD3 acts as a transcription factor for the gene PLOD2, which is crucial for the development of HP. The aim of the present work was to investigate the influence of miR-146a on the mechanism of increased collagen synthesis in the context of age-related arterial stiffening. The results showed that miR-146a is significantly upregulated in both old murine aortas and on rigid cell culture plates. The genes SMAD3 and PLOD2 were significantly down-regulated after transfection of miR-146a mimic and significantly up-regulated after transfection of miR-146a inhibitor. However, the results of the experiment with siRNA for SMAD3 were not significant. The Scar-in-a-jar assay showed that the miR-146a mimic decreased the synthesis of collagen and the miR-146a inhibitor increased the synthesis. The present results indicate that miR-146a has a protective effect on the development of vascular fibrosis. The regulation could take place via the TGF-β1/SMAD3 signaling pathway and via the increased formation of crosslinks by LH2 and its gene PLOD2. The collected data show for the first time that miR-146a as an important regulator of vascular fibrosis in the context of age-related arterial stiffening. MiR-146a is therefore a promising target for novel miR-based therapeutic approaches to arterial stiffness.2022-10-1

"Little Boxes (In the Archives)" (Parody)

A re-working of the Pete Seeger classic, "Little Boxes" (yes, I know it was written by Malvina Reynolds, but I'm only familiar with the Seeger version) for the amusement, hopefully, of my friends in the archives and information science community. I've been playing guitar for about a year, and I humbly welcome any constructive criticism or suggestions. I also seem to have managed to cut the top of my head off. I'm sure I'll get better at it