scholarly journals Ionizable lipid nanoparticles for in utero mRNA delivery

2021 ◽  
Vol 7 (3) ◽  
pp. eaba1028
Author(s):  
Rachel S. Riley ◽  
Meghana V. Kashyap ◽  
Margaret M. Billingsley ◽  
Brandon White ◽  
Mohamad-Gabriel Alameh ◽  
...  
Keyword(s):  
Messenger Rna ◽  
Gene Editing ◽  
Genetic Diseases ◽  
Lipid Nanoparticles ◽  
In Utero ◽  
Luciferase Mrna ◽  
Fetal Size ◽  
Mouse Fetuses ◽  
Clinical Advances ◽  
Immature Immune System

Clinical advances enable the prenatal diagnosis of genetic diseases that are candidates for gene and enzyme therapies such as messenger RNA (mRNA)–mediated protein replacement. Prenatal mRNA therapies can treat disease before the onset of irreversible pathology with high therapeutic efficacy and safety due to the small fetal size, immature immune system, and abundance of progenitor cells. However, the development of nonviral platforms for prenatal delivery is nascent. We developed a library of ionizable lipid nanoparticles (LNPs) for in utero mRNA delivery to mouse fetuses. We screened LNPs for luciferase mRNA delivery and identified formulations that accumulate within fetal livers, lungs, and intestines with higher efficiency and safety compared to benchmark delivery systems, DLin-MC3-DMA and jetPEI. We demonstrate that LNPs can deliver mRNAs to induce hepatic production of therapeutic secreted proteins. These LNPs may provide a platform for in utero mRNA delivery for protein replacement and gene editing.

scholarly journals Intracellular Routing and Recognition of Lipid-Based mRNA Nanoparticles

Pharmaceutics ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 945
Author(s):  
Christophe Delehedde ◽  
Luc Even ◽  
Patrick Midoux ◽  
Chantal Pichon ◽  
Federico Perche
Keyword(s):  
Gene Therapy ◽  
Immune Responses ◽  
Transient Expression ◽  
Messenger Rna ◽  
Lipid Nanoparticles ◽  
Delivery Systems ◽  
Clinical Applications ◽  
Cellular Proteins ◽  
Mrna Sequence ◽  
Cytoplasmic Expression

Messenger RNA (mRNA) is being extensively used in gene therapy and vaccination due to its safety over DNA, in the following ways: its lack of integration risk, cytoplasmic expression, and transient expression compatible with fine regulations. However, clinical applications of mRNA are limited by its fast degradation by nucleases, and the activation of detrimental immune responses. Advances in mRNA applications, with the recent approval of COVID-19 vaccines, were fueled by optimization of the mRNA sequence and the development of mRNA delivery systems. Although delivery systems and mRNA sequence optimization have been abundantly reviewed, understanding of the intracellular processing of mRNA is mandatory to improve its applications. We will focus on lipid nanoparticles (LNPs) as they are the most advanced nanocarriers for the delivery of mRNA. Here, we will review how mRNA therapeutic potency can be affected by its interactions with cellular proteins and intracellular distribution.

scholarly journals Pediatric and Adolescent Oncofertility in Male Patients—From Alpha to Omega

Genes ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 701
Author(s):  
Ovidiu Bîcă ◽  
Ioan Sârbu ◽  
Carmen Iulia Ciongradi
Keyword(s):  
Fertility Preservation ◽  
Gene Editing ◽  
Genetic Diseases ◽  
Reproductive Potential ◽  
Molecular Techniques ◽  
Male Population ◽  
Male Survivors ◽  
Male Patients ◽  
Survivors Of Childhood Cancer

This article reviews the latest information about preserving reproductive potential that can offer enhanced prospects for future conception in the pediatric male population with cancer, whose fertility is threatened because of the gonadotoxic effects of chemotherapy and radiation. An estimated 400,000 children and adolescents aged 0–19 years will be diagnosed with cancer each year. Fertility is compromised in one-third of adult male survivors of childhood cancer. We present the latest approaches and techniques for fertility preservation, starting with fertility preservation counselling, a clinical practice guideline used around the world and finishing with recent advances in basic science and translational research. Improving strategies for the maturation of germ cells in vitro combined with new molecular techniques for gene editing could be the next scientific keystone to eradicate genetic diseases such as cancer related mutations in the offspring of cancer survivors.

scholarly journals CRISPR Gene-Editing Models Geared Toward Therapy for Hereditary and Developmental Neurological Disorders

2021 ◽  
Vol 9 ◽  
Author(s):  
Poh Kuan Wong ◽  
Fook Choe Cheah ◽  
Saiful Effendi Syafruddin ◽  
M. Aiman Mohtar ◽  
Norazrina Azmi ◽  
...  
Keyword(s):  
Genetic Variation ◽  
Neurological Disorders ◽  
Gene Editing ◽  
Fragile X ◽  
Genetic Diseases ◽  
Basic Principles ◽  
Short Palindromic Repeat

Hereditary or developmental neurological disorders (HNDs or DNDs) affect the quality of life and contribute to the high mortality rates among neonates. Most HNDs are incurable, and the search for new and effective treatments is hampered by challenges peculiar to the human brain, which is guarded by the near-impervious blood-brain barrier. Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR), a gene-editing tool repurposed from bacterial defense systems against viruses, has been touted by some as a panacea for genetic diseases. CRISPR has expedited the research into HNDs, enabling the generation of in vitro and in vivo models to simulate the changes in human physiology caused by genetic variation. In this review, we describe the basic principles and workings of CRISPR and the modifications that have been made to broaden its applications. Then, we review important CRISPR-based studies that have opened new doors to the treatment of HNDs such as fragile X syndrome and Down syndrome. We also discuss how CRISPR can be used to generate research models to examine the effects of genetic variation and caffeine therapy on the developing brain. Several drawbacks of CRISPR may preclude its use at the clinics, particularly the vulnerability of neuronal cells to the adverse effect of gene editing, and the inefficiency of CRISPR delivery into the brain. In concluding the review, we offer some suggestions for enhancing the gene-editing efficacy of CRISPR and how it may be morphed into safe and effective therapy for HNDs and other brain disorders.

The Ethical Challenges of Gene Editing

2017 ◽  
Vol 42 (12) ◽  
pp. 3-4
Author(s):  
Jozef Zalot ◽  
Tadeusz Pacholczyk ◽  
Keyword(s):  
Catholic Church ◽  
Gene Editing ◽  
Human Life ◽  
Genetic Diseases ◽  
Future Generations ◽  
Human Embryos ◽  
Human Beings ◽  
Ethical Challenges ◽  
Oregon Health ◽  
Life Threatening

In August 2017, researchers at the Oregon Health and Science University announced that they had successfully used a gene editing technique known as CRISPR-Cas9 to repair disease-causing genes in human embryos. Some members of the scientific and medical communities have hailed the development as a way to ensure that life- threatening diseases are not passed on to future generations. But is gene editing always a good thing? The Catholic Church encourages scientific research that is ethical and serves the human good. In the future, CRISPR may be used to treat people with serious genetic diseases, such as hemophilia and sickle-cell anemia. However, for research on human beings to be ethical, it must be strictly therapeutic and must respect the dignity and sacredness of human life. Gene-editing techniques raise profound ethical challenges in both respects.

scholarly journals In utero gene editing for monogenic lung disease

2019 ◽  
Vol 11 (488) ◽  
pp. eaav8375 ◽  
Author(s):  
Deepthi Alapati ◽  
William J. Zacharias ◽  
Heather A. Hartman ◽  
Avery C. Rossidis ◽  
John D. Stratigis ◽  
...  
Keyword(s):  
Lung Disease ◽  
Lung Diseases ◽  
Gene Editing ◽  
In Utero ◽  
Lung Damage ◽  
Secretory Cells ◽  
Specific Gene ◽  
Alveolar Type ◽  
Diffuse Parenchymal Lung Disease ◽  
Parenchymal Lung

Monogenic lung diseases that are caused by mutations in surfactant genes of the pulmonary epithelium are marked by perinatal lethal respiratory failure or chronic diffuse parenchymal lung disease with few therapeutic options. Using a CRISPR fluorescent reporter system, we demonstrate that precisely timed in utero intra-amniotic delivery of CRISPR-Cas9 gene editing reagents during fetal development results in targeted and specific gene editing in fetal lungs. Pulmonary epithelial cells are predominantly targeted in this approach, with alveolar type 1, alveolar type 2, and airway secretory cells exhibiting high and persistent gene editing. We then used this in utero technique to evaluate a therapeutic approach to reduce the severity of the lethal interstitial lung disease observed in a mouse model of the human SFTPCI73T mutation. Embryonic expression of SftpcI73T alleles is characterized by severe diffuse parenchymal lung damage and rapid demise of mutant mice at birth. After in utero CRISPR-Cas9–mediated inactivation of the mutant SftpcI73T gene, fetuses and postnatal mice showed improved lung morphology and increased survival. These proof-of-concept studies demonstrate that in utero gene editing is a promising approach for treatment and rescue of monogenic lung diseases that are lethal at birth.

scholarly journals Aerosolizable Lipid Nanoparticles for Pulmonary Delivery of mRNA through Design of Experiments

Pharmaceutics ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 1042
Author(s):  
Hairui Zhang ◽  
Jasmim Leal ◽  
Melissa R. Soto ◽  
Hugh D. C. Smyth ◽  
Debadyuti Ghosh
Keyword(s):  
Design Of Experiments ◽  
Protein Expression ◽  
Messenger Rna ◽  
Pulmonary Delivery ◽  
Lipid Nanoparticles ◽  
In Vivo Studies ◽  
Molar Ratio ◽  
Mouse Lung ◽  
Intracellular Protein ◽  
Before And After

Messenger RNA is a class of promising nucleic acid therapeutics to treat a variety of diseases, including genetic diseases. The development of a stable and efficacious mRNA pulmonary delivery system would enable high therapeutic concentrations locally in the lungs to improve efficacy and limit potential toxicities. In this study, we employed a Design of Experiments (DOE) strategy to screen a library of lipid nanoparticle compositions to identify formulations possessing high potency both before and after aerosolization. Lipid nanoparticles (LNPs) showed stable physicochemical properties for at least 14 days of storage at 4 °C, and most formulations exhibited high encapsulation efficiencies greater than 80%. Generally, upon nebulization, LNP formulations showed increased particle size and decreased encapsulation efficiencies. An increasing molar ratio of poly-(ethylene) glycol (PEG)-lipid significantly decreased size but also intracellular protein expression of mRNA. We identified four formulations possessing higher intracellular protein expression ability in vitro even after aerosolization which were then assessed in in vivo studies. It was found that luciferase protein was predominately expressed in the mouse lung for the four lead formulations before and after nebulization. This study demonstrated that LNPs hold promise to be applied for aerosolization-mediated pulmonary mRNA delivery.

scholarly journals Nanomedicines to Deliver mRNA: State of the Art and Future Perspectives

Nanomaterials ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 364 ◽  
Author(s):  
Itziar Gómez-Aguado ◽  
Julen Rodríguez-Castejón ◽  
Mónica Vicente-Pascual ◽  
Alicia Rodríguez-Gascón ◽  
María Ángeles Solinís ◽  
...  
Keyword(s):  
Immune Responses ◽  
Plasmid Dna ◽  
Messenger Rna ◽  
Insertional Mutagenesis ◽  
Gene Editing ◽  
State Of The Art ◽  
Translational Efficiency ◽  
Future Perspectives ◽  
Therapeutic Tool

The use of messenger RNA (mRNA) in gene therapy is increasing in recent years, due to its unique features compared to plasmid DNA: Transient expression, no need to enter into the nucleus and no risk of insertional mutagenesis. Nevertheless, the clinical application of mRNA as a therapeutic tool is limited by its instability and ability to activate immune responses; hence, mRNA chemical modifications together with the design of suitable vehicles result essential. This manuscript includes a revision of the strategies employed to enhance in vitro transcribed (IVT) mRNA functionality and efficacy, including the optimization of its stability and translational efficiency, as well as the regulation of its immunostimulatory properties. An overview of the nanosystems designed to protect the mRNA and to overcome the intra and extracellular barriers for successful delivery is also included. Finally, the present and future applications of mRNA nanomedicines for immunization against infectious diseases and cancer, protein replacement, gene editing, and regenerative medicine are highlighted.

A New Era for Rare Genetic Diseases: Messenger RNA Therapy

2019 ◽  
Vol 30 (10) ◽  
pp. 1180-1189 ◽  
Author(s):  
Paolo G.V. Martini ◽  
Lin T. Guey
Keyword(s):  
Messenger Rna ◽  
Genetic Diseases ◽  
New Era ◽  
Rare Genetic Diseases

scholarly journals Cell-Selective Regulation of CFTR Gene Expression: Relevance to Gene Editing Therapeutics

Genes ◽  
2019 ◽  
Vol 10 (3) ◽  
pp. 235 ◽  
Author(s):  
Hannah Swahn ◽  
Ann Harris
Keyword(s):  
Gene Expression ◽  
Cystic Fibrosis ◽  
Gene Editing ◽  
Genetic Diseases ◽  
3D Structure ◽  
Three Dimensional ◽  
Cell Types ◽  
Regulatory Elements ◽  
Cftr Gene ◽  
Specific Expression

The cystic fibrosis transmembrane conductance regulator (CFTR) gene is an attractive target for gene editing approaches, which may yield novel therapeutic approaches for genetic diseases such as cystic fibrosis (CF). However, for gene editing to be effective, aspects of the three-dimensional (3D) structure and cis-regulatory elements governing the dynamic expression of CFTR need to be considered. In this review, we focus on the higher order chromatin organization required for normal CFTR locus function, together with the complex mechanisms controlling expression of the gene in different cell types impaired by CF pathology. Across all cells, the CFTR locus is organized into an invariant topologically associated domain (TAD) established by the architectural proteins CCCTC-binding factor (CTCF) and cohesin complex. Additional insulator elements within the TAD also recruit these factors. Although the CFTR promoter is required for basal levels of expression, cis-regulatory elements (CREs) in intergenic and intronic regions are crucial for cell-specific and temporal coordination of CFTR transcription. These CREs are recruited to the promoter through chromatin looping mechanisms and enhance cell-type-specific expression. These features of the CFTR locus should be considered when designing gene-editing approaches, since failure to recognize their importance may disrupt gene expression and reduce the efficacy of therapies.

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