Podcasts about oligonucleotide

Guidance Recap Podcast | Clinical Pharmacology Considerations for the Development of Oligonucleotide Therapeutics

In this new ACT ToxChats© episode, Dr. Jeffrey Foy, Vice President of Toxicology at Pepgen and Dr. Scott Henry, Vice President of Nonclinical Development at Ionis Pharmaceuticals are interviewed. Drs. Foy and Henry discuss the past, present, and future of oligonucleotides. Their focus is discussing the development and challenges that oligonucleotides face as a class of drugs.

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.02.23.529798v1?rss=1 Authors: Watanabe, N., Tone, Y., Nagata, T., Masuda, S., Saito, T., Motohashi, N., Takagaki, K., Aoki, Y., Takeda, S. Abstract: Exon-skipping therapy mediated by antisense oligonucleotides (ASOs) is expected to provide a therapeutic option for Duchenne muscular dystrophy (DMD). ASOs for exon skipping reported so far target a single continuous sequence in or around the target exon. In the present study, we investigated ASOs for exon 44 skipping (applicable to approximately 6% of all DMD patients) to improve activity by using a novel ASO design incorporating two connected sequences. Phosphorodiamidate morpholino oligomers targeting two separate sequences in exon 44 were created to simultaneously target two splicing regulators in exon 44, and their exon 44 skipping was measured. NS-089/NCNP-02 showed the highest skipping activity among the oligomers. NS-089/NCNP-02 also induced exon 44 skipping and dystrophin protein expression in cells from a DMD patient to whom exon 44 skipping is applicable. We also assessed the in vivo activity of NS-089/NCNP-02 by intravenous administration to cynomolgus monkeys. NS-089/NCNP-02 induced exon 44 skipping in skeletal and cardiac muscle of cynomolgus monkeys. In conclusion, NS-089/NCNP-02, an ASO with a novel connected-sequence design, showed both in vitro and in vivo exon-skipping activity. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2023.02.14.528473v1?rss=1 Authors: Mortberg, M. A., Gentile, J. E., Nadaf, N., Vanderburg, C., Simmons, S., Dubinsky, D., Slamin, A., Maldonado, S., Petersen, C. L., Jones, N., Kordasiewicz, H. B., Zhao, H. T., Vallabh, S. M., Minikel, E. V. Abstract: Antisense oligonucleotides (ASOs) dosed into cerebrospinal fluid (CSF) distribute broadly throughout the brain and hold the promise of treating myriad brain diseases by modulating RNA. CNS tissue is not routinely biopsied in living individuals, leading to reliance on CSF biomarkers to inform on drug target engagement. Animal models can link CSF biomarkers to brain parenchyma, but our understanding of how individual cells contribute to bulk tissue signal is limited. Here we employed single nucleus transcriptomics on tissue from mice treated with RNase H1 ASOs against Prnp and Malat1 and macaques treated with an ASO against PRNP. Activity was observed in every cell type, though sometimes with substantial differences in magnitude. Single cell RNA count distributions implied target suppression in every single sequenced cell, rather than intense knockdown in only some cells. Duration of action up to 12 weeks post-dose differed across cell types, being shorter in microglia than in neurons. Suppression in neurons was generally similar to, or more robust than, the bulk tissue. In macaques, PrP in CSF was lowered 40% in conjunction with PRNP knockdown across all cell types including neurons, arguing that a CSF biomarker readout is likely to reflect disease-relevant cells in a neuronal disorder. Copy rights belong to original authors. Visit the link for more info Podcast created by Paper Player, LLC

PepGen, a Cambridge, MA-based biotech, is advancing oligonucleotide therapeutics with the goal of transforming the treatment of severe neuromuscular and neurologic diseases has focused on delivery. SVP Head of Clinical Development Dr. Michelle Mellion explains PepGen's trial of PGN-EDO51, the company's lead product candidate for the treatment of DMD patients whose mutations are amenable to an exon 51 skipping approach. She also explains how the delivery of oligonucleotide therapies differ from other techniques in genetic medicine, and much more.

In this episode, we had the pleasure of speaking with Dr Rakesh N. Veedu on the topic of oligonucleotide therapeutics. If you're drawing a blank, we bet you've heard of the scientific pursuits into finding cures for HIV infection, macular degeneration, and spinal muscular atrophy. What about the endeavour to protect humanity against Covid? That one's pretty fresh and don't worry, we don't have divisive discussion around whether or not the vaccine is effective in the context of the current climate. We simply talk facts and get into the nitty gritty of how scientists manufacture DNA and RNA in order to diagnose and treat conditions beyond our comprehension. It's fascinating and I hope you'll join me in the conversation with DR Rakesh. Drop your notes on your preferred channels or find us on social media or via email at theperthbusinesspodcast@gmail.com. --- Send in a voice message: https://anchor.fm/perth-business-podcast/message

Oligonucleotide therapies can target the root cause of many diseases through the modulation of RNA expression and processing. Despite the promise of these medicines, their development has been limited by delivery challenges because they are not able to adequately reach heart and skeletal muscle, the critical affected tissues in neuromuscular diseases. PepGen is advancing next-generation oligonucleotide therapeutics that leverage its delivery platform technology to produce cell-penetrating peptide conjugates that improve the activity and tolerability of oligonucleotide therapies. We spoke to James McArthur, president and CEO of PepGen, about the company's platform technology for conjugating peptides with oligonucleotides, how this allows it to target hard to reach tissue, and why it opens the potential for new therapies to treat neuromuscular and other diseases.

Among the more prevalent genetic conditions, Duchenne muscular dystrophy  affects an estimated one in 3,500 male births worldwide.  It's caused by mutations of the DMD gene, which  regulates the production of a protein called dystrophin. Approved DMD treatments haven't demonstrated strong clinical outcomes, but James McArthur, Ph.D. and his team at PepGen are seeking to change that with a pipeline of disease-modifying peptide-conjugated oligonucleotide candidates derived from the company's Enhanced Delivery Oligonucleotide platform. The Business of Biotech caught up with Dr. McArthur at PepGen's Cambridge headquarters to learn more. Catch up on more gene therapy development advances on bioprocessonline.com.

Welcome to this week's episode of the Physician's Weekly podcast. I am your host, Dr. Rachel Giles, from Medicom Medical Publishers, in collaboration with Physician's Weekly.  Today we have two great interviews, one with the senior author on a recent trial using a unique medical approach published in Nature Medicine, and the other with our regular expert, who goes by the pseudonym Dr. MedLaw.But first, we talk with Professor Stephen Russell, from the University of Iowa Institute for Vision Research, Iowa City, IA, about how his team has been using oligonucleotide injections into the eyes of vision-impaired patients with Leber's congenital amaurosis, to force the transcription machinery to skip the exon bearing a mutation in a gene called CEP290. The trial has some surprising results, which were recently published in Nature Medicine. Enjoy listening!Further readingRussell SR, Drack AV, Cideciyan AV, Jacobson SG, Leroy BP, Van Cauwenbergh C, Ho AC, Dumitrescu AV, Han IC, Martin M, Pfeifer WL, Sohn EH, Walshire J, Garafalo AV, Krishnan AK, Powers CA, Sumaroka A, Roman AJ, Vanhonsebrouck E, Jones E, Nerinckx F, De Zaeytijd J, Collin RWJ, Hoyng C, Adamson P, Cheetham ME, Schwartz MR, den Hollander W, Asmus F, Platenburg G, Rodman D, Girach A. Intravitreal antisense oligonucleotide sepofarsen in Leber congenital amaurosis type 10: a phase 1b/2 trial. Nat Med. 2022 May;28(5):1014-1021. doi: 10.1038/s41591-022-01755-w. Epub 2022 Apr 4. PMID: 35379979; PMCID: PMC9117145.

Dr. Robert Brown discusses the Nature Medicine article, "Suppression of Mutant C9orf72 Expression by a Potent Mixed Backbone Antisense Oligonucleotide". Show references: https://www.nature.com/articles/s41591-021-01557-6

Dr. Jeff Ratliff discusses the suppression of mutant C9orf72 expression by a potent mixed backbone antisense oligonucleotide with Drs. Jonathan Watts and Robert Brown.

We discuss the different types of antisense oligonucleotide (AON/ASO) technologies and how these can be used therapeutically for FSHD. --- Send in a voice message: https://anchor.fm/peter-l-jones/message

Epilepsy research lead (PI) and lecturer in pharmacy, Cristina Reschke tells us about her work in both her exciting research into how microRNAs and circadian rhythm disruptions lead to epileptogenesis and seizures and educating pharmacists of the future regarding the epilepsies.---More about Cristina:  https://www.torierobinson.com/epilepsy-sparks-insights/cristina-reschke---Glossary:There are many terms used in this recording which you can find in the Epilepsy Sparks Glossary: https://www.epilepsysparks.com/glossary---Follow Torie on:- Twitter: https://twitter.com/torierobinson10- LinkedIn: https://www.linkedin.com/torierobinson- Instagram: https://www.instagram.com/torierobinson10- Facebook: https://www.facebook.com/TorieRobinsonSpeaker- Check out the website: https://www.torierobinson.com

I sat down  with Cia in April to talk about her great love Regulatory RNA!We discussed ..What is an #oligonucleotide, and more specifically #ASO, antisense oligo?Are regulatory RNAs abundant in living cells?How do you commonly purify such molecules?Do modifications of the oligos affect the #purification?You can get more info atBlog: https://lnkd.in/gFaYNzgLinkedIn pageWebsite www.bio-works.comYou can also watch over 180 interviews and product demos with experts across Drug Discovery, Development and Manufacturing on my YouTube channel RSK Life Sciences Media here  https://www.youtube.com/channel/UC9vdET2xLlGweqveW6JmQdQ

Go online to PeerView.com/DYM860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, an expert in neurology discusses the use of antisense oligonucleotide therapies in patients with amyotrophic lateral sclerosis (ALS). Upon completion of this accredited CE activity, participants should be better able to: Identify common genes and cellular processes implicated in ALS pathogenesis, Recognize the mechanisms of action, efficacy, and safety profiles of emerging ALS therapies including antisense oligonucleotides, Discuss how the ALS treatment paradigm may change as a result of emerging therapies, Educate patients with ALS about ongoing clinical trials evaluating novel disease-modifying therapies.

Go online to PeerView.com/DYM860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, an expert in neurology discusses the use of antisense oligonucleotide therapies in patients with amyotrophic lateral sclerosis (ALS). Upon completion of this accredited CE activity, participants should be better able to: Identify common genes and cellular processes implicated in ALS pathogenesis, Recognize the mechanisms of action, efficacy, and safety profiles of emerging ALS therapies including antisense oligonucleotides, Discuss how the ALS treatment paradigm may change as a result of emerging therapies, Educate patients with ALS about ongoing clinical trials evaluating novel disease-modifying therapies.

Go online to PeerView.com/DYM860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, an expert in neurology discusses the use of antisense oligonucleotide therapies in patients with amyotrophic lateral sclerosis (ALS). Upon completion of this accredited CE activity, participants should be better able to: Identify common genes and cellular processes implicated in ALS pathogenesis, Recognize the mechanisms of action, efficacy, and safety profiles of emerging ALS therapies including antisense oligonucleotides, Discuss how the ALS treatment paradigm may change as a result of emerging therapies, Educate patients with ALS about ongoing clinical trials evaluating novel disease-modifying therapies.

Go online to PeerView.com/DYM860 to view the activity, download slides and practice aids, and complete the post-test to earn credit. In this activity, an expert in neurology discusses the use of antisense oligonucleotide therapies in patients with amyotrophic lateral sclerosis (ALS). Upon completion of this accredited CE activity, participants should be better able to: Identify common genes and cellular processes implicated in ALS pathogenesis, Recognize the mechanisms of action, efficacy, and safety profiles of emerging ALS therapies including antisense oligonucleotides, Discuss how the ALS treatment paradigm may change as a result of emerging therapies, Educate patients with ALS about ongoing clinical trials evaluating novel disease-modifying therapies.

Learn about the RLR gene editing tool; Victorian-era emoji; and why “night shift” screen settings don't help you sleep. RLR is a new gene editing tool that's safer and more useful than CRISPR by Cameron Duke Move over CRISPR, the retrons are coming. (2021). EurekAlert! https://www.eurekalert.org/pub_releases/2021-04/wifb-moc042921.php  Schubert, M. G., Goodman, D. B., Wannier, T. M., Kaur, D., Farzadfard, F., Lu, T. K., Shipman, S. L., & Church, G. M. (2021). High-throughput functional variant screens via in vivo production of single-stranded DNA. Proceedings of the National Academy of Sciences, 118(18), e2018181118. https://doi.org/10.1073/pnas.2018181118  Swingle, B., Markel, E., & Cartinhour, S. (2010). Oligonucleotide recombination: A hidden treasure. Bioengineered Bugs, 1(4), 265–268. https://doi.org/10.4161/bbug.1.4.12098  Irving, M. (2021, May 4). Harvard gene-editing tool “sneaks” DNA into cells without making cuts. New Atlas. https://newatlas.com/biology/retron-library-recombineering-gene-editing-crispr/  ‌Are Retrons the Next CRISPR? | Cell And Molecular Biology. (2021). Labroots. https://www.labroots.com/trending/cell-and-molecular-biology/20347/retrons-crispr  Flowers Were Emoji For Victorians by Anna Todd Romie Stott. (2016, August 15). How Flower-Obsessed Victorians Encoded Messages in Bouquets. Atlas Obscura; Atlas Obscura. https://www.atlasobscura.com/articles/how-flowerobsessed-victorians-encoded-messages-in-bouquets  The meaning of flowers, Floriography, language of flowers, advice hints and tips from All Florists.co.uk. Allflorists.co.uk. http://www.allflorists.co.uk/advice_flowerMeanings.asp  "Night shift" screen settings don't really help you sleep by Kelsey Donk Is night shift really helping you sleep better? (2021). EurekAlert! https://www.eurekalert.org/pub_releases/2021-04/byu-ins042621.php Duraccio, K. M., Zaugg, K. K., Blackburn, R. C., & Jensen, C. D. (2021). Does iPhone night shift mitigate negative effects of smartphone use on sleep outcomes in emerging adults? Sleep Health. https://doi.org/10.1016/j.sleh.2021.03.005  Follow Curiosity Daily on your favorite podcast app to learn something new every day withCody Gough andAshley Hamer — for free!  See omnystudio.com/listener for privacy information.

As interest in oligonucleotide therapeutics increases, there is an ever-growing need for analytical techniques that can properly analyse and quantitate these complex biomolecules. Mass spectrometry (MS) is required to analyse impurities, degradants, and other biological/chemical modifications and is the method of choice for the analysis of synthetic oligonucleotides. Sixfold Bioscience, in collaboration with the National Measurement Laboratory (NML) have been using the Agilent AdvancedBio 6545XT LC/Q-TOF to develop a method to analyse complex oligonucleotides of various lengths with volatile ion pair reagents. This method is used for quality control of oligonucleotides that are subsequently assembled into Mergos™ for use in in vivo applications. In addition to the analytical capabilities currently being utilised by Sixfold, the Agilent 1290 Infinity II Preparative system is also being used for oligonucleotide purification. Both reverse-phase (C18) and anion-exchange (IEX) chromatography is regularly carried out to assist in Sixfold's purification workflow. As a result of this work, and as an extension to the work currently being carried out by the Tate group at Imperial College, a Metabolomics group is in the process of being formed at the Molecular Sciences Research Hub. This group will aim to solve challenges related to the analysis of biomolecules and will take advantage of the combined know-how and expertise of scientists at Imperial College, Sixfold Bioscience and Agilent. Dr James Rushworth, Research Associate at Sixfold Bioscience. For Research Use Only. Not for use in diagnostic procedures. DE44320.2788078704

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.11.12.379164v1?rss=1 Authors: Burbano, L. E., Li, M., Jancovsky, N., Jafar-Nejad, P., Richards, K., Sedo, A., Soriano, A., Rollo, B., Jia, L., Gazina, E., Piltz, S., Adikusuma, F., Thomas, P. Q., Rigo, F., Reid, C. A., Maljevic, S., Petrou, S. Abstract: Developmental and epileptic encephalopathies (DEE) are characterized by pharmacoresistant seizures with concomitant intellectual disability. Epilepsy of infancy with migrating focal seizures (EIMFS) is one of the most severe of these syndromes. De novo mutations in ion channels, including gain-of-function variants in KCNT1, have been found to play a major role in the etiology of EIMFS. Here, we test a potential precision therapeutic approach in KCNT1-associated DEE using a gene silencing antisense oligonucleotide (ASO) approach. The homozygous p.P924L (L/L) mouse model recapitulates the frequent, debilitating seizures and developmental compromise that are seen in patients. After a single intracerebroventricular bolus injection of a Kcnt1 gapmer ASO in symptomatic mice at postnatal day 40, seizure frequency was significantly reduced, behavioral abnormalities improved, and overall survival was extended compared to mice treated with a control ASO (non-hybridizing sequence). ASO administration at neonatal age was also well-tolerated and effective in controlling seizures and extending the lifespan of treated animals. The data presented here provides a proof of concept for ASO-based gene silencing as a promising therapeutic approach in KCNT1-associated epilepsies. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.09.09.289900v1?rss=1 Authors: Li, M., Jancovski, N., Jafar-nejad, P., Burbano, L., Rollo, B., Richards, K., Drew, L., Sedo, A., pachernegg, S., Soriano, A., Jia, L., Blackburn, T., Roberts, B., Nemiroff, A., Dalby, K., Maljevic, S., Reid, C., Rigo, F., Petrou, S. Abstract: The clinical spectrum associated with SCN2A de novo mutations (DNMs) continues to expand and includes autism spectrum disorder with or without seizures, in addition to early and late seizure onset developmental and epileptic encephalopathies (DEEs). Recent biophysical studies on SCN2A variants suggest that the majority of early seizure onset DEE DNMs cause gain of function. Gain of function in SCN2A, the principal sodium channel of excitatory pyramidal neurons, would result in heightened neuronal activity and is likely to underlie the pathology seen in early seizure onset DEE patients. Supratherapeutic dosing of the non-selective sodium channel blocker phenytoin, is effective in controlling seizures in these patients but does not impact neurodevelopment, raising the idea that more profound and specific reduction in SCN2A function could significantly improve clinical outcome. To test the potential therapeutic benefit of reducing SCN2A in early seizure onset DEE we centrally administered an antisense oligonucleotide (ASO) targeting mouse Scn2a (Scn2a ASO) to a mouse model of human SCN2A early seizure onset DEE. Mice were genetically engineered to harbour the human equivalent SCN2A p.R1882Q mutation (Q/+), one of the most recurrent mutations in early seizure onset DEE. Q/+ mice presented with spontaneous seizures at postnatal day (P) 1 and did not survive beyond P30. Intracerebroventricular Scn2a ASO administration into Q/+ mice between P1-2 (that reduced Scn2a mRNA levels by 50%) significantly extended lifespan and markedly reduced spontaneous seizures occurrence. Across a range of cognitive and motor behavioural tests, Scn2a ASO treated Q/+ mice were largely indistinguishable from wildtype (+/+) mice. Further improvements in survival and behaviour were seen by adjustment of dosing regimens during development. Scn2a ASO efficacy was also evident at the cellular level. Whole cell patch clamp recording showed that Scn2a ASO administration reversed changes in neuronal excitability in layer 2/3 pyramidal neurons of Q/+ mice to levels seen in +/+ mice. Safety was assessed in +/+ mice and showed a developmental stage dependent tolerability and a favourable therapeutic index. This study suggests that a human SCN2A gapmer ASO could profoundly and safely impact early seizure onset DEE patients and heralds a new era of precision therapy in neurodevelopmental disorders. Copy rights belong to original authors. Visit the link for more info

Link to bioRxiv paper: http://biorxiv.org/cgi/content/short/2020.07.31.216721v1?rss=1 Authors: Rigo, F., Jafar-nejad, P., Powers, B., Soriano, A., Zhao, H., Norris, D. A., Matson, J., DeBrosse-Serra, B., Watson, J., Narayanan, P., Mazur, C., Kordasiewicz, H., Swayze, E. E., Narayanan, P. Abstract: Antisense oligonucleotides (ASOs) have emerged as a new class of drugs to treat a wide range of diseases, including neurological indications. Spinraza, an ASO that modulates splicing of SMN2 RNA, has shown profound disease modifying effects in Spinal Muscular Atrophy (SMA) patients, energizing the field to develop ASOs for other neurological disorders. While SMA specifically affects spinal motor neurons, other disorders affect different central nervous system (CNS) regions, neuronal, and non-neuronal cells. Therefore, it is critically important to characterize ASO distribution and activity in all major CNS structures and cell types to have a better understanding of which neurological diseases are amenable to ASO therapy. Here we present for the first time the atlas of ASO distribution and activity in the CNS of mice, rats, and non-human primates (NHP), species commonly used in preclinical therapeutic development. Following central administration of an ASO to rodents, we observe widespread distribution and robust activity throughout the CNS in neurons, oligodendrocytes, astrocytes, and microglia. This is also the case in NHP, despite larger CNS volume and more complex neuroarchitecture. Our results demonstrate that ASO drugs are well suited for treating a wide range of neurological diseases for which no effective treatments are available. Copy rights belong to original authors. Visit the link for more info

Dr. Crowell interviews Dr. Timothy Yu about his article "Patient-customized oligonucleotide therapy for a rare genetic disease." Read the NEJM article: https://www.nejm.org/doi/full/10.1056/NEJMoa1813279. 

David Giljohann likens his company’s spherical nucleic acid (SNA) constructs to Koosh balls. But instead of having elastic strings extending from their core, this new class of immunomodulatory and gene regulating drugs are covered with nucleic acids that stick out in all directions. The arrangement allows these oligonucleotides to be taken up by all cell types and can be delivered throughout the body allowing them to overcome some of the limitations of existing nucleic acid therapies. We spoke to Giljohann, CEO of Exicure, about the company’s SNA therapies, how they work, and why they have the potential to treat a wide range of conditions including rare neurological diseases.

Dr. Jason Crowell discusses Richard Robinson's article on an antisense oligonucleotide therapy for a rare variant of Batten Disease. To learn more, read the article "An Antisense Oligonucleotide Therapy Looks Promising for a Rare Form of Batten Disease" by Robinson in the June 20, 2019, issue of Neurology Today—available online at http://bit.ly/NT-Batten.

Dr. Arthur A. Levin, the interviewee, is the Executive Vice President of Research and Development at Avidity Biosciences. Stephen Morrissey, the interviewer, is the Executive Managing Editor of the Journal. A.A. Levin. Treating Disease at the RNA Level with Oligonucleotides. N Engl J Med 2019;380:57-70.

Dr. Sabrina Leslie, Associate Professor of Physics and Quantitative Life Sciences at McGill University, leads listeners on an interesting journey into the complex and important world of molecule imaging and research. As an experienced researcher and respected university leader at McGill, Dr. Leslie founded her single-molecule imaging laboratory in 2012. The lab was instrumental in the development of novel techniques to discover and explore complex interactions and dynamics of macromolecules. Dr. Leslie has pursued doctoral studies in optical physics, has completed extensive research on biomolecules in liquids, and explored her passion for imaging in a health sciences context. She is a recipient of a Mary Fieser Postdoctoral Fellowship, and her work at Harvard led to the invention of a single-molecule imaging technology known as Convex Lens-induced Confinement (CLiC). And it was this groundbreaking work in the field that thrust her onto the international stage as a noted pioneer in single-molecule investigations with a broad range of applications. Dr. Leslie earned her Ph.D. in Optical and Atomic Physics at the University of California at Berkeley Dr. Leslie discusses her lab's intensive work deconstructing molecular interactions to answer questions in biology and develop new drugs for medical and therapeutic applications. She talks about the importance of time scales, speed of camera, and resolution, in their work as they seek to visualize the interactions in real time. By using cameras they are able to watch molecules for a long time and deconstruct the methods of operation. Collaborating with biochemists and cancer scientists, as well as applied mathematicians, the Leslie Lab's single-molecule investigations of protein and DNA interactions, have provided significant new information about gene regulation mechanisms. Specifically, Dr. Leslie's work has also been focused on exploring a wide range of interactions and dynamics of proteins and DNA, as well as other macromolecules, and with regard to molecular shape and environment.  The macromolecule researcher details her lab's groundbreaking work with the imaging technology known as Convex Lens-induced Confinement (CLiC). As she explains, the process works by essentially squeezing the tiny molecules into little traps so that the essence of the molecule is completely held in focus, viewable from all angles. And by utilizing fluorescent labels they are able to closely track them for observation.  She details some of their current research regarding DNA binding with a focus on applications and therapeutics, etc. As they continued their experimentation they began introducing other molecules and polymers, to fill up space and then observe how DNA probes find their target sites. As she states, they have a particular interest in the nexus between the physics and the biology, where structural elements can have an impact on the biochemical reactions. Further, she talks about some surprising discoveries regarding proteins and droplet formation simulation.  In concert with biochemists in Montreal, Dr. Leslie's team is instigating CLiC studies of non-membranous organelles (NMOs). Liquid droplets form when proteins and nucleic acids phase-separate from cellular material, similar to how oil separates from water. Importantly, there is significant evidence suggesting that misregulation of MMOs is associated with select diseases such as Alzheimer's. Lastly, Dr. Leslie expounds upon some of the areas of research they are most excited to dig into, such as gene editing, and she gives an overview of their collaborations with pharmaceutical companies on oligonucleotide therapeutics. (Oligonucleotide therapeutics is an inventive class of biopharmaceutical products composed of short strings of synthetic nucleotides that resemble the actual building blocks for DNA.)

Oligonucleotide therapeutics are all medications that utilize RNA- or DNA-based techniques as the effective entity. The nonclinical development of these therapeutics presents unique challenges and these are discussed here with Drs. Sven Korte from Covance, Brian Vuillemenot from Genentech and Scott Henry from Ionis Pharmaceuticals, all experts in oligo therapeutic development.Disclaimer: The views and opinions expressed in this podcast are those of the guest speakers and do not necessarily reflect the official policy or position of the American College of Toxicology. For the College’s policy on the ethical treatment of animals in research, please see our comprehensive policy statement at www.actox.org.

CHI recently interviewed Dr. Dmitry Samarsky, Senior Vice President of Technology and Global Business Development at OliX Pharmaceuticals, regarding the evolution of nucleic acid-based therapeutics; challenges within this space; recent advances in chemistry and delivery; and the greatest opportunities within this space over the next few years. Discussion questions include: 1 Given the obvious benefit of oligonucleotide therapies being able to dramatically expand target space, why haven’t they impacted the drug development landscape as much as Biologics or small Molecules? What challenges have been encountered within this field? 2 It seems as if there has be a resurgence of interest in the discovery and development of oligonucleotide therapeutics. Why do you think this is? What has changed over the past few years? 3 You are hosting a workshop during the upcoming Oligonucleotide Therapeutics and Delivery conference, providing an overview of oligonucleotide therapeutics from discovery to manufacturing. Can you tell us more about this workshop? Who should attend this? What can they expect to learn during this? 4 Oligonucleotide therapeutics seem to be evolving rapidly, but in your opinion, where do you see the greatest opportunities within this space over the next few years? For more information, visit http://www.HealthTech.com/Oligonucleotide/

Ian C. Eperon, Department of Biochemistry, Henry Wellcome Building, Leicester - UNITED KINGDOM speaks on "Mechanisms, structures and complexes of a targeted oligonucleotide enhancer of SMN2 exon 7 splicing". ICGEB 2nd Post-EURASNET Symposium “RNA Alternative Splicing” This seminar has been recorded at ICTP Trieste by ICGEB Trieste

Cohen syndrome is a rare autosomal recessive disorder with a complex phenotype including psychomotor retardation, microcephaly, obesity with slender extremities, joint laxity, progressive chorioretinal dystrophy/myopia, intermittent isolated neutropenia, a cheerful disposition, and characteristic facial features. The COH1 gene, which contains 62 exons, is so far the only gene known to be associated with Cohen syndrome. Point mutations, deletions and duplications have been described in this gene. Oligonucleotide arrays have reached a resolution which allows the detection of intragenic deletions and duplications, especially in large genes such as COH1.

Guest: Dr. Annemieke Aartsma-Rus, DMD Genetic Therapy Group, Dept. of Human Genetics, Leiden University Medical Center, The Netherlands Access an abstract of this Month's Featured Research Article: Progress in therapeutic antisense applications for neuromuscular disorders. Eur J Hum Genet. 2010 Feb;18(2):146-53.

Guest: Dr. Annemieke Aartsma-Rus, DMD Genetic Therapy Group, Dept. of Human Genetics, Leiden University Medical Center, The Netherlands Access an abstract of this Month's Featured Research Article: Progress in therapeutic antisense applications for neuromuscular disorders. Eur J Hum Genet. 2010 Feb;18(2):146-53.

The present work follows a course of analytical description, formulation development, and practical application of (NF-κB decoy oligonucleotide-loaded) gelatin nanoparticles. The introduction of asymmetrical flow field-flow fractionation (AF4) in the analysis of colloidal drug carrier systems was exemplarily described for gelatin nanoparticles (CHAPTER I), stable freeze-dried formulations of empty and oligonucleotide loaded gelatin nanoparticles were successfully developed (CHAPTER II), and gelatin nanoparticles were proven as effective tool for the targeted delivery of an NF-κB decoy oligonucleotide to Kupffer cells within a hepatic ischemia reperfusion rat model (CHAPTER III).

Widerstandsarterien spielen eine wichtige Rolle bei der Durchblutungsregulation. Bisher konnte der wichtigste endotheliale Dilatator in diesen Gefäßen, EDHF, nicht eindeutig identifiziert werden, da pharmakologische Inhibitoren unspezifische Nebenwirkungen aufwiesen. Die spezifische Inhibition von Enzymen mittels Antisensetechnik konnte in intakten Arterien nicht durchgeführt werden, da diese nur über einen kurzen Zeitraum funktionell intakt erhalten werden konnten. Im Rahmen dieser Dissertation wurde ein neues Organkulturmodell entwickelt, in dem erstmalig die endothelabhängigen EDHF- und NO-vermittelten Dilatationen über 48 h vollständig erhalten werden konnten. Zusätzlich entwickelten die kultivierten Arterien einen mit dem frisch isolierter Arterien vergleichbaren Spontantonus und zeigten eine myogene Reaktion, die sich in Kinetik und Ausmaß der Kontraktion nicht von den Kontrollarterien unterschied. Ebenso kontrahierten die chronisch perfundierten Arterien auf Stimulation mit Noradrenalin und dilatierten nach Applikation des NO-Donors SNP in vergleichbarem Ausmaß wie frisch isolierte Arterien. Um zu untersuchen, ob möglicherweise eine CytochromP450-Epoxygenase in der Signalkaskade des EDHF eine Rolle spielt, wurde zunächst die Expression von CYP2C8 in Widerstandsarterien mittels rtPCR und in-situ-Hybridisierung nachgewiesen. Da mit dem Organkulturmodell die Arterien funktionell vollständig intakt gehalten werden konnten, wurde die Wirkung von Antisense-Oligonucleotiden, die gegen CYP2C8 gerichtet waren, untersucht. Mittels konfokaler Mikroskopie konnte gezeigt werden, dass die FITC-markierten Oligonucleotide sich nur in der Intima befanden und die Transfektion des Endothels eine hohe Effizienz aufwies. Die Transfektion hatte keinen Effekt auf die NA-induzierte Kontraktion, auf die durch NS1619 (KCa-Kanalöffner)- oder die SNP- vermittelte Relaxation, was zeigt, dass die Funktion des glatten Muskels durch die Transfektion unbeeinträchtigt blieb. Die EDHF-vermittelten Dilatationen wurden durch die Transfektion mit den Antisense-Oligonucleotiden um 76% und die korrespondierenden Calciumabfälle um 58 % reduziert, während die Kontrolltransfektionen mit Scrambled- oder Senseoligonucleotiden keinen Einfluss auf die EDHF-mediierten Dilatationen hatten. Die endothelialen Calciumanstiege nach Stimulation mit ACh blieben in den Antisense-transfizierten Arterien unverändert. Das bedeutet, dass die Signaltransduktion der ACh-Rezeptoren durch die Transfektion funktionell nicht beeinträchtigt wurde. Auf diese Weise konnte mit einem spezifischen Inhibitor gezeigt werden, dass CYP2C8 eine EDHF-Synthase ist oder dessen Metabolit einen permissiven Faktor für einen anderen EDHF darstellt und ein elementarer Bestandteil der EDHF-Signalkaskade ist. Zusätzlich wurden mit diesem Organkulturmodell die Auswirkungen des kardiovaskulären Risikofaktors Hochdruck durch isolierte Erhöhung des transmuralen Drucks auf 120 und 160 mmHg (SMA120 bzw. SMA160) während einer Kulturperiode (48 h) untersucht. In den funktionellen Testungen zeigten sich nach 48 h geringere Außendurchmesserwerte in SMA120 und SMA160 im Sinne eines Remodelings. Der erhöhte Perfusionsdruck führte darüber hinaus zu einer Verstärkung der Noradrenalin-vermittelten Kontraktion. Dies ist jedoch nicht durch eine Erhöhung der Calciumsensitivität der Myofilamente zu erklären, da diese im Vergleich zur Kontrolle unverändert war, sondern durch eine Verstärkung der NA-induzierten Calciumanstiege. Neben den Veränderungen in der glatten Muskulatur zeigte sich insbesondere auch eine Beeinträchtigung der Endothel-vermittelten Relaxationen. Die NO-mediierte Dilatation wurde durch die chronische Perfusion bei 120 mmHg um 38% reduziert und bei SMA160 vollständig aufgehoben. Ebenso wurde die EDHF-vermittelte Relaxation bei SMA120 um 20 % und bei SMA160 um 47% verringert und der korrespondierende Calciumabfall um 41 % reduziert. Diese Reduktion der endothelialen Dilatationen wurde nicht durch eine Erhöhung der Elastance der Arterienwand hervorgerufen, da die dosisabhängige SNP-mediierte Relaxation unbeeinträchtigt war. Zusätzlich scheint eine strukturelle Schädigung des Endothels durch den erhöhten Druck unwahrscheinlich, da mittels Rasterelektronenmikroskopie keine Schäden an der Intima dargestellt werden konnten. Die Expression des ACh-Rezeptors scheint auch nicht in dem Maße verringert zu sein, dass sich daraus die verringerten NO- und EDHF-mediierten Relaxationen erklären ließen, da der endotheliale Calciumanstieg in SMA120 im Vergleich zu SMA45 unverändert war. Daher wird die Beeinträchtigung durch den erhöhten Druck in einem nachgeschalteten Signaltransduktionsweg vermutet. Erhöhter transmuraler Druck hat in diesem Modell innerhalb von 2 Tagen schon zu einer erheblichen Beeinträchtigung der endothelialen Funktionen und zu einer verstärkten Reaktivität des glatten Muskels in Widerstandsarterien geführt. Zwar ist eine Erhöhung des transmuralen Drucks für 48 h nicht mit einem jahrelang bestehenden Hypertonus vergleichbar, jedoch könnte man die so erhobenen Befunde als Hinweis werten, dass eine frühzeitige konsequente antihypertensive Therapie sinnvoll ist, um die druckinduzierte Verstärkung der glattmuskulären Reaktivität und die Einschränkung der Endothelfunktion zu verringern und eine daraus resultierende weitere Erhöhung des Blutdruckes zu verhindern.

Non-isotopic in situ hybridization of chromosome-specific alphoid DNA probes has become a potent tool in the study of numerical aberrations of specific human chromosomes at all stages of the cell cycle. In this paper, we describe approaches for the rapid generation of such probes using the polymerase chain reaction (PCR), and demonstrate their chromosome specificity by fluorescence in situ hybridization to normal human metaphase spreads and interphase nuclei. Oligonucleotide primers for conserved regions of the alpha satellite monomer were used to generate chromosome-specific DNA probes from somatic hybrid cells containing various human chromosomes, and from DNA libraries from sorted human chromosomes. Oligonucleotide primers for chromosome-specific regions of the alpha satellite monomer were used to generate specific DNA probes for the pericentromeric heterochromatin of human chromosomes 1, 6, 7, 17 and X directly from human genomic DNA.