Delivery Of Genetic Information Research Articles

Tailor made: the art of therapeutic mRNA design.

mRNA medicine is a new and rapidly developing field in which the delivery of genetic information in the form of mRNA is used to direct therapeutic protein production in humans. This approach, which allows for the quick and efficient identification and optimization of drug candidates for both large populations and individual patients, has the potential to revolutionize the way we prevent andtreat disease. A key feature of mRNA medicines is their high degree of designability, although the design choices involved are complex. Maximizing the production of therapeutic proteins from mRNA medicines requires a thoroughunderstanding of how nucleotide sequence, nucleotide modification and RNA structure interplay to affect translational efficiency and mRNA stability. In this Review, we describe the principles that underlie the physical stability and biological activity of mRNA and emphasize their relevance to the myriad considerations that factor into therapeutic mRNA design.

Role of Stem Cell-Derived Microvesicles in Cardiovascular Disease.

The role of stem cells in augmenting reparative processes in the heart after ischemic injury has been successfully demonstrated in small and large animal models. However, the outcomes of cell therapy in clinical trials have been somewhat variable, with overall effects of autologous stem cell therapies demonstrating a modest improvement in cardiac structure and function. How stem cells repair the heart after cardiac injury is still not well understood. Most recent studies suggest that adult derived stem cells act primarily through paracrine signaling to exert beneficial effects, including modulation of immune response, stimulation of new blood vessel formation, or by inducing mature myocytes to transiently reenter the cell cycle, rather than robust direct differentiation of the transplanted cells into myocytes. In addition, data from multiple laboratory results confirmed clearance of stem cells themselves within a few days still leading to functional benefits further confirming the role of paracrine signaling in augmenting cardiac reparative processes rather than direct differentiation of cells. These findings rapidly evolved the field of extracellular vesicles specifically microvesicles (MVs) as they are active hubs of autocrine, paracrine, and endocrine signaling targeting different biological processes. The beneficial effects seen after stem cell transplantation could be linked to the cardioprotective factors packaged in the MVs secreted from stem cells. Therefore, stem cell MVs provide a new avenue for the treatment of cardiovascular disease through a multitude of mechanisms including cellular communication within the stem cell niches, delivery of genetic information, regulation of the immune system in the heart, and stimulation of angiogenesis which will be discussed in this review.

MAGENTA (Making Genetic testing accessible): a prospective randomized controlled trial comparing online genetic education and telephone genetic counseling for hereditary cancer genetic testing

BackgroundStudies have consistently indicated that the majority of individuals meeting the US Prevention Services Task Force guidelines for genetic testing have not had genetic counseling or testing. Despite increased availability and lower costs of multiplex cancer gene panels, there remains a gap in genetics services that has not been addressed by the current care delivery models. Lower cost of DNA sequencing with online patient-initiated ordering could increase test availability, but the ideal quantity and delivery method of patient education is not known. We hypothesized that online genetic education and testing with access to board certified genetic counselors could improve access to genetic testing while maintaining test quality and clinical utility. The MAGENTA (MAking GENetic Testing Accessible) trial is a nationwide randomized study designed to compare the effectiveness of online genetic education with pre- and post-test telephone genetic counseling to three potentially more accessible alternative approaches: online genetic education with optional telephone counseling, online genetic education with required pre-test telephone genetic counseling, and online genetic education with required post-test telephone genetic counseling.Methods3000 women nationwide will undergo genetic testing for 19 hereditary cancer genes. This is a randomized four-arm non-inferiority study with equal randomization. The four study arms were selected to independently assess the delivery of genetic information both before and after genetic testing (pre-test and post-test) by either requiring telephone genetic counseling or providing only online education with optional telephone counseling. Patients have post-test telephone counseling when testing positive for a pathogenic inherited mutation in all four arms. Surveys measuring psychological, behavioral and cognitive state are completed online at baseline, 3 months, 12 months and 24 months post-results disclosure. The primary study outcome is cancer-risk distress at 3 months post-result disclosure.DiscussionThis trial will assess the use of a genetic service model using online access and electronic education, while evaluating the need for personal pre- and post-test genetic counseling. Data from this study may lead to increased options for delivery of genetic testing and possibly increase access to genetic testing. Identifying more individuals with inherited cancer susceptibility will allow targeted cancer prevention.Trial registrationClinicaltrials.gov: NCT02993068 (registered December 14, 2016).

MRNA-basierte Therapien für eine passive Immunisierung

The delivery of genetic information to enable the body to produce its own tailored medicine is a promising alternative to recombinant protein therapies. As such, the administration of mRNA offers a potent, flexible and apparently safe platform. Here we discuss the principles and perspectives of mRNA-based therapies focusing on its application in the field of passive immunization in general and for infectious diseases in particular.

MRNA mediates passive vaccination against infectious agents, toxins, and tumors.

The delivery of genetic information has emerged as a valid therapeutic approach. Various reports have demonstrated that mRNA, besides its remarkable potential as vaccine, can also promote expression without inducing an adverse immune response against the encoded protein. In the current study, we set out to explore whether our technology based on chemically unmodified mRNA is suitable for passive immunization. To this end, various antibodies using different designs were expressed and characterized in vitro and in vivo in the fields of viral infections, toxin exposure, and cancer immunotherapies. Single injections of mRNA–lipid nanoparticle (LNP) were sufficient to establish rapid, strong, and long‐lasting serum antibody titers in vivo, thereby enabling both prophylactic and therapeutic protection against lethal rabies infection or botulinum intoxication. Moreover, therapeutic mRNA‐mediated antibody expression allowed mice to survive an otherwise lethal tumor challenge. In conclusion, the present study demonstrates the utility of formulated mRNA as a potent novel technology for passive immunization.

NEI's Audacious Goals Initiative

In 2013, the National Eye Institute (NEI) launched a program with an unconventional goal. The NEI challenged the ophthalmic and vision research community to seek creative and pioneering initiatives that would fundamentally advance research over the next 10 to 15 years. The response was overwhelming, with >500 researchers and clinicians submitting ideas. The NEI centered the initiative around well-defined prizes rather than grants, drawing in the expertise and perspectives of people not typically involved in vision research. Both physicians and research scientists were among the winners and almost half of submissions came from people who had never received funding from the National Institutes of Health. In addition to harnessing new perspectives, the initiative’s fresh approach removed the constraints that go hand in hand with grant writing and allowed more room for imagination. Researchers did not have to worry about being incremental and safe in their ideas in order to ensure funding, and could instead be ambitious and truly audacious. A total of 474 eligible applications were winnowed down to 81 by clinicians and scientists, who provided input to a panel of judges through numerical scores, letter grades, and technical comments. Using the same judging methods as the technical advisors, a federal judging panel selected 10 winning entries. Next, in February 2013, the NEI convened a meeting of international thought leaders in vision research. The 10 top ideas had scientist leaders representing the ideas as discussion facilitators, and scientists from vastly different backgrounds met in different rooms at the NEI with the facilitators to discuss these 10 ideas. The process itself was audacious and transformative, with farreaching discussions and reports from each session. The invited scientists narrowed the 10winners into 3 high-priority areas of research. The 10 winning ideas (Table 1) are characterized by their relevance, boldness, feasibility, farreaching scope, and potential to be transformative. The NEI senior staff consulted with the National Advisory Eye Council to identify a single audacious goal: Regenerating neurons and neural connections in the eye and visual system. Two research areas were also designated as high priority: Molecular therapy for eye disease and intersection of aging and biological mechanisms of eye disease. The first area of high-priority research, molecular therapy for eye disease, focuses on the development of treatments through the modification and delivery of genetic information. The past several years have seen the advent of new technologies for precise gene correction in vivo and the unraveling of the genetic underpinning of many eye diseases, transforming this idea from a far-off vision to a realistic goal. Although the idea of performing treatments at the molecular level has driven research on blinding eye

Gene Therapy for Prostate Cancer

Cancer remains a leading cause of morbidity and mortality. Despite advances in understanding, detection, and treatment, it accounts for almost one-fourth of all deaths per year in Western countries. Prostate cancer is currently the most commonly diagnosed noncutaneous cancer in men in Europe and the United States, accounting for 15% of all cancers in men. As life expectancy of individuals increases, it is expected that there will also be an increase in the incidence and mortality of prostate cancer. Prostate cancer may be inoperable at initial presentation, unresponsive to chemotherapy and radiotherapy, or recur following appropriate treatment. At the time of presentation, patients may already have metastases in their tissues. Preventing tumor recurrence requires systemic therapy; however, current modalities are limited by toxicity or lack of efficacy. For patients with such metastatic cancers, the development of alternative therapies is essential. Gene therapy is a realistic prospect for the treatment of prostate and other cancers, and involves the delivery of genetic information to the patient to facilitate the production of therapeutic proteins. Therapeutics can act directly (eg, by inducing tumor cells to produce cytotoxic agents) or indirectly by upregulating the immune system to efficiently target tumor cells or by destroying the tumor's vasculature. However, technological difficulties must be addressed before an efficient and safe gene medicine is achieved (primarily by developing a means of delivering genes to the target cells or tissue safely and efficiently). A wealth of research has been carried out over the past 20 years, involving various strategies for the treatment of prostate cancer at preclinical and clinical trial levels. The therapeutic efficacy observed with many of these approaches in patients indicates that these treatment modalities will serve as an important component of urological malignancy treatment in the clinic, either in isolation or in combination with current approaches.

DNA therapeutics: a feasible option for treatment of inflammatory diseases?

Recombinant proteins can be very potent, but their therapeutic application can be strongly hampered by inappropriate distribution, dosage, kinetics or toxic effects. Targeted delivery of proteins such as cytokines would be strongly desirable. DNA therapeutics comprise the delivery of genetic information on a piece of DNA as therapeutic prodrug. This prodrug can be transcribed and translated into a protein (the actual drug) within the target cells, preferably in a tissue-specific and bio-regulated fashion. Basically, two types of nonviral gene transfer systems [1] have been developed: particle-based systems, with DNA packaged into cationic lipids or polymers; and physical techniques which are based on combining DNA with a physical device. Intramuscular administration of naked DNA has already proven as interesting concept for vaccination [2], despite the low efficiency of the method. Two physical device technologies, electroporation and the gene gun, were found to enhance gene expression levels up to 1000- fold over injection of naked DNA alone. This enhancement has also recently been shown by several groups to trigger immune responses against defined antigens in several species [3]. We have generated particle-based systems that can target gene delivery and expression into distant target tissues. We use DNA polyplexes conjugated with cell-binding ligands such as transferrin for receptor-mediated endocytosis. The surface charge of complexes is masked by covalent coating with polyethylenglycol (PEG). Tumor targeting has been demonstrated in mouse models after systemic administration. With systemically applied tumor necrosis factor (TNF) alpha gene, tumor necrosis and regression of tumors was observed, but no systemic TNF-related side effects. Opportunities to apply local or systemic DNA therapeutics for inflammatory diseases will be discussed.