Optimization of lipid nanoparticles for gene editing of the liver via intraduodenal delivery.

Lipid nanoparticles (LNPs) have recently emerged as successful gene delivery platforms for a diverse array of disease treatments. Efforts to optimize their design for common administration methods such as intravenous injection, intramuscular injection, or inhalation, revolve primarily around the addition of targeting ligands or the choice of ionizable lipid. Here, we employed a multi-step screening method to optimize the type of helper lipid and component ratios in a plasmid DNA (pDNA) LNP library to efficiently deliver pDNA through intraduodenal delivery as an indicative route for oral administration. By addressing different physiological barriers in a stepwise manner, we down-selected effective LNP candidates from a library of over 1000 formulations. Beyond reporter protein expression, we assessed the efficiency in non-viral gene editing in mouse liver mediated by LNPs to knockdown PCSK9 and ANGPTL3 expression, thereby lowering low-density lipoprotein (LDL) cholesterol levels. Utilizing an all-in-one pDNA construct with Strep. pyogenes Cas9 and gRNAs, our results showcased that intraduodenal administration of selected LNPs facilitated targeted gene knockdown in the liver, resulting in a 27% reduction in the serum LDL cholesterol level. This LNP-based all-in-one pDNA-mediated gene editing strategy highlights its potential as an oral therapeutic approach for hypercholesterolemia, opening up new possibilities for DNA-based gene medicine applications.

Machine Learning Elucidates Design Features of Plasmid DNA Lipid Nanoparticles for Cell Type-Preferential Transfection.

For cell and gene therapies to become more broadly accessible, it is critical to develop and optimize non-viral cell type-preferential gene carriers such as lipid nanoparticles (LNPs). Despite the effectiveness of high throughput screening (HTS) approaches in expediting LNP discovery, they are often costly, labor-intensive, and often do not provide actionable LNP design rules that focus screening efforts on the most relevant chemical and formulation parameters. Here we employed a machine learning (ML) workflow using well-curated plasmid DNA LNP transfection datasets across six cell types to maximize chemical insights from HTS studies and has achieved predictions with 5-9% error on average depending on cell type. By applying Shapley additive explanations to our ML models, we unveiled composition-function relationships dictating cell type-preferential LNP transfection efficiency. Notably, we identified consistent LNP composition parameters that enhance in vitro transfection efficiency across diverse cell types, such as ionizable to helper lipid ratios near 1:1 or 10:1 and the incorporation of cationic/zwitterionic helper lipids. In addition, several parameters were found to modulate cell type-preferentiality, including the ionizable and helper lipid total molar percentage, N/P ratio, cholesterol to PEGylated lipid ratio, and the chemical identity of the helper lipid. This study leverages HTS of compositionally diverse LNP libraries and ML analysis to understand the interactions between lipid components in LNP formulations; and offers fundamental insights that contribute to the establishment of unique sets of LNP compositions tailored for cell type-preferential transfection.

Extending traditional antibody therapies: Novel discoveries in immunotherapy and clinical applications.

Immunotherapy has been well regarded as one of the safer and antigen-specific anti-cancer treatments compared to first-generation chemotherapy. Since Coley's discovery, researchers focused on engineering novel antibody-based therapies. Including artificial and modified antibodies, such as antibody fragments, antibody-drug conjugates, and synthetic mimetics, the variety of immunotherapy has been rapidly expanding in the last few decades. Genetic and chemical modifications to monoclonal antibody have been brought into academia, in vivo trials, and clinical applications. Here, we have looked around antibodies overall. First, we elucidate the antibody structure and its cytotoxicity mechanisms. Second, types of therapeutic antibodies are presented. Additionally, there is a summarized list of US Food and Drug Administration (FDA)-approved therapeutic antibodies and recent clinical trials. This review provides a comprehensive overview of both the general function of therapeutic antibodies and a few main variations in development, including recent advent with the proposed mechanism of actions, and we introduce types of therapeutic antibodies, clinical trials, and approved commercial immunotherapeutic drugs.

Longitudinal trajectories of ketamine use among young injection drug users.

BACKGROUND: Ketamine is a dissociative anaesthetic that became increasingly popular in the club and rave scene in the 1980s and 1990s. Reports surfaced in the late 1990s indicating that ketamine was being injected in several U.S. cities by young injection drug users (IDUs). Since all studies on ketamine injection were cross-sectional, a longitudinal study was undertaken in 2005 to determine: characteristics of young IDUs who continue to inject ketamine; frequency of ketamine injection over an extended time period; risks associated with ongoing ketamine injection; and environmental factors that impact patterns of ketamine use. METHODS: Young IDUs aged 16-29 with a history of injecting ketamine (n=101) were recruited from public locations in Los Angeles and followed during a 2-year longitudinal study. A semi-structured instrument captured quantitative and qualitative data on patterns of ketamine injection and other drug use. A statistical model sorted IDUs who completed three or more interviews (n=66) into three groups based upon patterns of ketamine injection at baseline and follow-up. Qualitative analysis focused on detailed case studies within each group. RESULTS: IDUs recruited at baseline were typically in their early 20s, male, heterosexual, white, and homeless. Longitudinal injection trajectories included: "Moderates," who injected ketamine several times per year (n=5); "Occasionals," who injected ketamine approximately once per year (n=21); and "Abstainers," who did not inject any ketamine during follow-up (n=40). Findings suggest that ketamine is infrequently injected compared to other drugs such as heroin, cocaine, and methamphetamine. Most IDUs who begin injecting ketamine will stop or curb use due to: negative or ambivalent experiences associated with ketamine; an inability to find the drug due to declining supply; or maturing out of injecting drugs more generally. CONCLUSION: Reducing ketamine injection among young IDUs may best be accomplished by targeting particular groups of IDUs identified in this study, such as homeless youth and homeless travellers.

THE FIRST INJECTION EVENT: DIFFERENCES AMONG HEROIN, METHAMPHETAMINE, COCAINE, AND KETAMINE INITIATES.

This article describes how the drug type injected at the first injection event is related to characteristics of the initiate, risk behaviors at initiation, and future drug-using trajectories. A diverse sample (n=222) of young injection drug users (IDUs) were recruited from public settings in New York, New Orleans, and Los Angeles during 2004 and 2005. The sample was between 16 and 29 years old, and had injected ketamine at least once in the preceding two years. Interview data was analyzed both quantitatively and qualitatively. Young IDUs initiated with four primary drug types: heroin (48.6%), methamphetamine (20.3%), ketamine (17.1%), and cocaine (14%). Several variables evidenced statistically significant relationships with drug type: age at injection initiation, level of education, region of initiation, setting, mode of administration, patterns of self-injection, number of drugs ever injected, current housing status, and their hepatitis C virus (HCV) status. Qualitative analyses revealed that rationale for injection initiation and subjective experiences at first injection differed by drug type.