Molecular mechanisms of amyotrophic lateral sclerosis as broad therapeutic targets for gene therapy applications utilizing adeno-associated viral vectors

Despite the devastating clinical outcome of the neurodegenerative disease, amyotrophic lateral sclerosis (ALS), its etiology remains mysterious. Approximately 90% of ALS is characterized as sporadic, signifying that the patient has no family history of the disease. The development of an impactful disease modifying therapy across the ALS spectrum has remained out of grasp, largely due to the poorly understood mechanisms of disease onset and progression. Currently, ALS is invariably fatal and rapidly progressive. It is hypothesized that multiple factors can lead to the development of ALS, however, treatments are often focused on targeting specific familial forms of the disease (10% of total cases). There is a strong need to develop disease modifying treatments for ALS that can be effective across the full ALS spectrum of familial and sporadic cases. Although the onset of disease varies significantly between patients, there are general disease mechanisms and progressions that can be seen broadly across ALS patients. Therefore, this review explores the targeting of these widespread disease mechanisms as possible areas for therapeutic intervention to treat ALS broadly. In particular, this review will focus on targeting mechanisms of defective protein homeostasis and RNA processing, which are both increasingly recognized as design principles of ALS pathogenesis. Additionally, this review will explore the benefits of gene therapy as an approach to treating ALS, specifically focusing on the use of adeno-associated virus (AAV) as a vector for gene delivery to the CNS and recent advances in the field

The future of recombinant host defense peptides

The antimicrobial resistance crisis calls for the discovery and production of new antimicrobials. Host defense peptides (HDPs) are small proteins with potent antibacterial and immunomodulatory activities that are attractive for translational applications, with several already under clinical trials. Traditionally, antimicrobial peptides have been produced by chemical synthesis, which is expensive and requires the use of toxic reagents, hindering the large-scale development of HDPs. Alternatively, HDPs can be produced recombinantly to overcome these limitations. Their antimicrobial nature, however, can make them toxic to the hosts of recombinant production. In this review we explore the different strategies that are used to fine-tune their activities, bioengineer them, and optimize the recombinant production of HDPs in various cell factories.info:eu-repo/semantics/publishedVersio

Modular feedback keyboard

International audienceThe modular feedback keyboard that we are going to describe here was brought out in october 1988 and its construction is the result of research of many years in our laboratory on the instrumental relationship within the framework of real-time sound synthesis and computer assisted musical creation. An international patent covers both its modularity principle as well as its motor technology, that was specially developed for the keyboard performance

Gene editing and Rett syndrome: does it make the cut?

Rett syndrome (RTT) is a rare neurogenetic disorder caused by pathogenic variants of the Methyl CpG binding protein 2 (MECP2) gene. The RTT is characterized by apparent normal early development followed by regression of communicative and fine motor skills. Comorbidities include epilepsy, severe cognitive impairment, and autonomic and motor dysfunction. Despite almost 60 clinical trials and the promise of a gene therapy, no cure has yet emerged with treatment remaining symptomatic. Advances in understanding RTT has provided insight into the complexity and exquisite control of MECP2 expression, where loss of expression leads to RTT and overexpression leads to MECP2 duplication syndrome. Therapy development requires regulated expression that matches the spatiotemporal endogenous expression of MECP2 in the brain. Gene editing has revolutionized gene therapy and promises an exciting strategy for many incurable monogenic disorders, including RTT, by editing the native locus and retaining endogenous gene expression. Here, we review the literature on the currently available editing technologies and discuss their limitations and applicability to the treatment of RTT

AAV-p40 Bioengineering Platform for Variant Selection Based on Transgene Expression

The power of AAV directed evolution for identifying novel vector variants with improved properties is well established, as evidenced by numerous publications reporting novel AAV variants. However, most capsid variants reported to date have been identified using either replication-competent selection platforms or PCR-based capsid DNA recovery methods, which can bias the selection towards efficient replication or unproductive intracellular trafficking, respectively. A central objective of this study was to validate a functional transduction (FT)-based method for rapid identification of novel AAV variants based on AAV capsid mRNA expression in target cells. We performed a comparison of the FT platform to existing replication competent strategies. Based on the selection kinetics and function of novel capsids identified in an in vivo screen in a xenograft model of human hepatocytes, we identified the mRNA-based FT selection as the most optimal AAV selection method. Lastly, to gain insight into the mRNA-based selection mechanism driven by the native AAV-p40 promoter, we studied its activity in a range of in vitro and in vivo targets. We found AAV-p40 to be a ubiquitously active promoter that can be modified for cell type-specific expression by incorporating binding sites for silencing transcription factors, allowing for cell-type-specific library selection

Operational Performance of MOSFIRE with Its Cryogenic Configurable Slitmask Unit at the W. M. Keck Observatory

The Multi-Object Spectrograph for Infrared Exploration (MOSFIRE) achieved first light on the W. M. Keck Observatory’s Keck I telescope on 4 April 2012 and quickly became the most popular Keck I instrument. One of the primary reasons for the instrument’s popularity is that it uses a configurable slitmask unit developed by the Centre Suisse d’Electronique et Microtechnique (CSEM SA) to isolate the light from up to 46 objects simultaneously. In collaboration with the instrument development team and CSEM engineers, the Keck observatory staff present how MOSFIRE is successfully used, and we identify what contributed to routine and trouble free nighttime operations

AAV capsid bioengineering in primary human retina models

Adeno-associated viral (AAV) vector-mediated retinal gene therapy is an active field of both pre-clinical as well as clinical research. As with other gene therapy clinical targets, novel bioengineered AAV variants developed by directed evolution or rational design to possess unique desirable properties, are entering retinal gene therapy translational programs. However, it is becoming increasingly evident that predictive preclinical models are required to develop and functionally validate these novel AAVs prior to clinical studies. To investigate if, and to what extent, primary retinal explant culture could be used for AAV capsid development, this study performed a large high-throughput screen of 51 existing AAV capsids in primary human retina explants and other models of the human retina. Furthermore, we applied transgene expression-based directed evolution to develop novel capsids for more efficient transduction of primary human retina cells and compared the top variants to the strongest existing benchmarks identified in the screening described above. A direct side-by-side comparison of the newly developed capsids in four different in vitro and ex vivo model systems of the human retina allowed us to identify novel AAV variants capable of high transgene expression in primary human retina cells

Operational Performance of MOSFIRE with Its Cryogenic Configurable Slitmask Unit at the W. M. Keck Observatory

The Multi-Object Spectrograph for Infrared Exploration (MOSFIRE) achieved first light on the W. M. Keck Observatory’s Keck I telescope on 4 April 2012 and quickly became the most popular Keck I instrument. One of the primary reasons for the instrument’s popularity is that it uses a configurable slitmask unit developed by the Centre Suisse d’Electronique et Microtechnique (CSEM SA) to isolate the light from up to 46 objects simultaneously. In collaboration with the instrument development team and CSEM engineers, the Keck observatory staff present how MOSFIRE is successfully used, and we identify what contributed to routine and trouble free nighttime operations

Assessment of pre-clinical liver models based on their ability to predict the liver-tropism of AAV vectors

The liver is a prime target for in vivo gene therapies using recombinant adeno-associated viral vectors (rAAV). Multiple clinical trials have been undertaken for this target in the past 15 years, however we are still to see market approval of the first liver-targeted AAV-based gene therapy. Inefficient expression of the therapeutic transgene, vector-induced liver toxicity and capsid, and/or transgene-mediated immune responses reported at high vector doses are the main challenges to date. One of the contributing factors to the insufficient clinical outcomes, despite highly encouraging preclinical data, is the lack of robust, biologically- and clinically-predictive preclinical models. To this end, this study reports findings of a functional evaluation of six AAV vectors in twelve preclinical models of the human liver, with the aim to uncover which combination of models is the most relevant for the identification of AAV capsid variant for safe and efficient transgene delivery to primary human hepatocytes. The results, generated by studies in models ranging from immortalized cells, iPSC-derived and primary hepatocytes, and primary human hepatic organoids to in vivo models, increased our understanding of the strengths and weaknesses of each system. This should allow the development of novel gene therapies targeting the human liver

Bogactwo i bieda: próba refleksji humanistycznej

Ze wstępu: "Niniejsza książka jest efektem pracy zespołu realizującego program badań „Bogactwo i bieda - postawy, opinie, zachowania (badania w środowisku studentów Akademii Górniczo- Hutniczej)” w ramach grantu uczelnianego (G. U. Z. 10.430). Projekt ten nawiązuje do tradycji badań środowiska studenckiego AGH, które zapoczątkował przed laty profesor Julian Bugiel. Prezentowana praca tworzy pewną całość wraz z wątkami problemowymi realizowanych wcześniej badań: w latach 2000-2001 „Agresja i przemoc - postawy, opinie, zachowania (badania w środowisku studentów AGH)” (G. U. Z. 21.10.430.69), jak również z badaniami przeprowadzonymi w roku 2002 „Tolerancja i nietolerancja - postawy, opinie, zachowania (badania w środowisku studentów AGH)” (G. U. Z. 21.10.430.48)."(...