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To predict whether preclinical lipid nanoparticle (LNP) delivery will translate in humans, it is necessary to understand whether the mechanism used by LNPs to enter cells is conserved across species. In mice, non-human primates, and humans, LNPs deliver RNA to hepatocytes by adsorbing apolipoprotein E (ApoE), which binds low-density lipoprotein receptor (LDLR). A growing number of LNPs can deliver RNA to nonhepatocytes, suggesting that ApoE- and LDLR-independent interactions could affect LNP tropism. To evaluate this hypothesis, we developed a universal DNA barcoding system that quantifies how chemically distinct LNPs deliver small interfering RNA in any mouse model, including genetic knockouts. We quantified how 98 different LNPs targeted 11 cell types in wildtype, LDLR

Less than 20% of the protein coding genome is thought to be targetable using small molecules. mRNA therapies are not limited in the same way since in theory, they can silence or edit any gene by encoding CRISPR nucleases, or alternatively, produce any missing protein. Yet not all mRNA therapies are equally likely to succeed. Over the past several years, an increasing number of clinical trials with siRNA- and antisense oligonucleotide-based drugs have revealed three key concepts that will likely extend to mRNA therapies delivered by nonviral systems. First, scientists have come to understand that some genes make better targets for RNA therapies than others. Second, scientists have learned that the type and position of chemical modifications made to an RNA drug can alter its therapeutic window, toxicity, and bioavailability. Third, scientists have found that safe and targeted drug delivery vehicles are required to ferry mRNA therapies into diseased cells. In this study, we apply these learnings to cystic fibrosis (CF). We also describe lessons learned from a subset of CF gene therapies that have already been tested in patients. Finally, we highlight the scientific advances that are still required for nonviral mRNA- or CRISPR-based drugs to treat CF successfully in patients.

Cells that were previously described as homogeneous are composed of subsets with distinct transcriptional states. However, it remains unclear whether this cell heterogeneity influences the efficiency with which lipid nanoparticles (LNPs) deliver messenger RNA therapies in vivo. To test the hypothesis that cell heterogeneity influences LNP-mediated mRNA delivery, we report here a new multiomic nanoparticle delivery system called single-cell nanoparticle targeting-sequencing (SENT-seq). SENT-seq quantifies how dozens of LNPs deliver DNA barcodes and mRNA into cells, the subsequent protein production and the transcriptome, with single-cell resolution. Using SENT-seq, we have identified cell subtypes that exhibit particularly high or low LNP uptake as well as genes associated with those subtypes. The data suggest that cell subsets have distinct responses to LNPs that may affect mRNA therapies.

PIP3, a bioactive lipid upstream of the PI3K/AKT/mTOR pathway, decreased protein translated from nanoparticle-delivered mRNA., The biological pathways that affect drug delivery in vivo remain poorly understood. We hypothesized that altering cell metabolism with phosphatidylinositol (3,4,5)-triphosphate (PIP3), a bioactive lipid upstream of the metabolic pathway PI3K (phosphatidylinositol 3-kinase)/AKT/ mTOR (mammalian target of rapamycin) would transiently increase protein translated by nanoparticle-delivered messenger RNA (mRNA) since these pathways increase growth and proliferation. Instead, we found that PIP3 blocked delivery of clinically-relevant lipid nanoparticles (LNPs) across multiple cell types in vitro and in vivo. PIP3-driven reductions in LNP delivery were not caused by toxicity, cell uptake, or endosomal escape. Interestingly, RNA sequencing and metabolomics analyses suggested an increase in basal metabolic rate. Higher transcriptional activity and mitochondrial expansion led us to formulate two competing hypotheses that explain the reductions in LNP-mediated mRNA delivery. First, PIP3 induced consumption of limited cellular resources, “drowning out” exogenously-delivered mRNA. Second, PIP3 triggers a catabolic response that leads to protein degradation and decreased translation.

Background Prucalopride, a selective, high-affinity 5-hydroxytryptamine 4 receptor agonist, stimulates gastrointestinal and colonic motility and alleviates common symptoms of chronic constipation (CC) in adults. The relative efficacy by gender has not been evaluated.

It is a descriptive study with the aim of studying the prevalence of tuberculosis in the State of Acre from 1995 to 2001. The data were collected from the forms of the Program for Tuberculosis Control of the Health and Sanitation Office. The data show that the incidence has been maintaining an average of 343.7 new cases a year. The pulmonary type was the most prevalent one, accounting for 90.0%. Among the age groups, the 20-49 age range, with 1347 (56.0%) and the 60 and over-60 age range, with 412 (17.0%), stand out. From 1998 to 2001 the male sex was the most attacked one by the disease, when the year 2000 accounted for 58.2% of the cases. The average of the cure rate was 71.0% and abandonment of treatment was 14.8% in 2001 and 32.3% in 1995. The smallest percentage of death occurred in 1995 (1.0%) and the largest one in 1997 (7.0%). Five cases were notified for the occurrence of coinfection between TB/AIDS and HIV/TB. The program's population coverage was within 90% every year. From the 22 cities, 6 (27.3%) still do not have any action for controlling the disease.

Trata-se de um estudo descritivo com o objetivo de estudar a prevalência da tuberculose no Estado do Acre, no período de 1995 a 2001. Os dados foram levantados junto aos formulários do Programa de Controle da Tuberculose da Secretaria de Saúde e Saneamento. Os dados mostram que a incidência tem mantido uma média de 343,7 casos novos por ano. A forma pulmonar foi a mais encontrada, representando 90,0%. Dentre as faixas etárias destacam-se a de 20-49 anos, com 1347 (56,0%) e a de 60 e mais anos com 412 (17,0%). No período de 1998 a 2001 o sexo masculino foi o mais acometido pela doença onde o ano de 2000 representou 58,2% dos casos. A média da taxa de cura foi de 71,0% e o abandono do tratamento com 14,8% em 2001 e 32,3% em 1995. Para o óbito o menor percentual foi em 1995 (1,0%) e o maior em 1997 (7,0%). Para a ocorrência de co-infecção entre TB/AIDS e HIV/TB foram notificados cinco casos. A cobertura populacional do programa em todos os anos esteve dentro da faixa de 90%, sendo que dos 22 municípios 6 (27,3%) ainda não possuem nenhuma ação de controle da doença. It is a descriptive study with the aim of studying the prevalence of tuberculosis in the State of Acre from 1995 to 2001. The data were collected from the forms of the Program for Tuberculosis Control of the Health and Sanitation Office. The data show that the incidence has been maintaining an average of 343.7 new cases a year. The pulmonary type was the most prevalent one, accounting for 90.0%. Among the age groups, the 20-49 age range, with 1,347 (56.0%) and the 60 and over-60 age range, with 412 (17.0%), stand out. From 1998 to 2001 the male sex was the most attacked one by the disease, when the year 2000 accounted for 58.2% of the cases. The average of the cure rate was 71.0% and abandonment of treatment was 14.8% in 2001 and 32.3% in 1995. The smallest percentage of death occurred in 1995 (1.0%) and the largest one in 1997 (7.0%). Five cases were notified for the occurrence of coinfection between TB/AIDS and HIV/TB. The program's population coverage was within 90% every year. From the 22 cities, 6 (27.3%) still do not have any action for controlling the disease. Se trata de un estudio descriptivo con el objetivo de estudiar la prevalencia de la tuberculosis en el Estado de Acre, en el período de 1995 a 2001. Los datos se levantaron junto a los formularios del Programa de Control de la Tuberculosis da Secretaría de Salud y Saneamiento. Los datos muestran que la incidencia ha mantenido un promedio de 343,7 casos nuevos por año. La forma pulmonar fue la más encontrada, representando 90,0%. Entre las edades se destacan la de 20-49 años, con 1347 (56,0%) y la de 60 y más años con 412 (17,0%). En el período de 1998 a 2001 el sexo masculino fue el más acometido por la enfermedad donde el año de 2000 representó 58,2% de los casos. El promedio de la tasa de cura fue de 71,0% y el abandono del tratamiento con 14,8% en 2001 y 32,3% en 1995. Para el óbito el menor porcentual fue en 1995 (1,0%) y el mayor en 1997 (7,0%). Para los casos de coinfección entre TB/SIDA y VHI/TB se notificaron cinco casos. La cobertura poblacional del programa en todos los años estuvo dentro de la franja de 90%, y de los 22 municipios 6 (27,3%) todavía no poseen ninguna acción de control de la enfermedad.

Background: Mesalazine (5-aminosalicylic acid) is standard first-line treatment for mild-to-moderate ulcerative colitis (UC), and has been studied for the treatment of diverticulitis (DV). Although there is some evidence of improved efficacy in induction of remission in UC with high-dose treatment, data are limited on the safety of long-term high-dose mesalazine treatment. This analysis examines the safety profile of highdose (4.8 g/d) MMX mesalazine administered for 2 years to patients with DV, pooled from 2 studies. Methods: Two Phase 3, randomized, double-blind, placebocontrolled, multicenter studies (PREVENT1, NCT00545740; PREVENT2, NCT00545103; sponsor, Shire Development LLC) evaluated the efficacy and safety of 3 dose regimens of oncedaily MMX mesalazine (1.2 g, 2.4 g, or 4.8 g) versus placebo in the prevention of recurrent DV in adult patients over a period of 104 wks. Results: A total of 1,182 total patients were enrolled in PREVENT1 and PREVENT2, and 301 were randomized to receive 4.8 g/d MMX mesalazine. Of these, 299 were administered 1 dose of MMX mesalazine (mean [SD] age, 55.6 [11.9] y; 46.8% male). Mean (SD) treatment exposure was 64.4 (42.7)wks. Among patients who received 1 dose of MMX mesalazine, 211 (70.6%) reported 1 treatment-emergent adverse event (TEAE; 1006 events reported); 59 (19.7%) patients experienced only mild TEAEs, 114 (38.1%) experienced TEAEs with a maximum severity of moderate, and 38 (12.7%) experienced severe TEAEs. Serious TEAEs were reported by 32 (10.7%) patients (38 events). A total of 51 (17.1%) patients experienced TEAEs determined to be related to study treatment (102 events), and 4 (1.3%) patients reported serious TEAEs related to study treatment (4 events). A total of 36 (12.0%) patients experienced TEAEs that led to withdrawal from their respective studies. The most frequently reported TEAEs (>5%) were abdominal pain (11.0%), diarrhoea (10.0%), headache (9.3%), back pain (6.3%), urinary tract infection (6.0%), nausea (5.3%), and upper respiratory tract infection (5.3%). Conclusions: Pooled analysis of adverse event data from longterm exposure to MMX mesalazine 4.8 g/d in patients with a history of DV showed that high-dose MMX mesalazine was welltolerated for 2 years.