An evaluation of oligonucleotide-based therapeutic strategies for polyQ diseases

<p>Abstract</p> <p>Background</p> <p>RNA interference (RNAi) and antisense strategies provide experimental therapeutic agents for numerous diseases, including polyglutamine (polyQ) disorders caused by CAG repeat expansion. We compared the potential of different oligonucleotide-based strategies for silencing the genes responsible for several polyQ diseases, including Huntington's disease and two spinocerebellar ataxias, type 1 and type 3. The strategies included nonallele-selective gene silencing, gene replacement, allele-selective SNP targeting and CAG repeat targeting.</p> <p>Results</p> <p>Using the patient-derived cell culture models of polyQ diseases, we tested various siRNAs, and antisense reagents and assessed their silencing efficiency and allele selectivity. We showed considerable allele discrimination by several SNP targeting siRNAs based on a weak G-G or G-U pairing with normal allele and strong G-C pairing with mutant allele at the site of RISC-induced cleavage. Among the CAG repeat targeting reagents the strongest allele discrimination is achieved by miRNA-like functioning reagents that bind to their targets and inhibit their translation without substantial target cleavage. Also, morpholino analog performs well in mutant and normal allele discrimination but its efficient delivery to cells at low effective concentration still remains a challenge.</p> <p>Conclusions</p> <p>Using three cellular models of polyQ diseases and the same experimental setup we directly compared the performance of different oligonucleotide-based treatment strategies that are currently under development. Based on the results obtained by us and others we discussed the advantages and drawbacks of these strategies considering them from several different perspectives. The strategy aimed at nonallele-selective inhibiting of causative gene expression by targeting specific sequence of the implicated gene is the easiest to implement but relevant benefits are still uncertain. The gene replacement strategy that combines the nonallele-selective gene silencing with the expression of the exogenous normal allele is a logical extension of the former and it deserves to be explored further. Both allele-selective RNAi approaches challenge cellular RNA interference machinery to show its ability to discriminate between similar sequences differing in either single base substitutions or repeated sequence length. Although both approaches perform well in allele discrimination most of our efforts are focused on repeat targeting due to its potentially higher universality.</p

Inflammatory Proteins HMGA2 and PRTN3 as Drivers of Vulvar Squamous Cell Carcinoma Progression.

Current knowledge on the biology of squamous cell vulvar carcinoma (VSCC) is limited. We aimed to identify protein markers of VSCC tumors that would permit to stratify patients by progression risk. Early-stage tumors from patients who progressed (progVSCC) and from those who were disease-free (d-fVSCC) during follow-up, along with normal vulvar tissues were examined by mass spectrometry-based proteomics. Differentially expressed proteins (DEPs) were then verified in solid tissues and blood samples of patients with VSCC tumors and vulvar premalignant lesions. In progVSCC vs. d-fVSCC tumors, the immune response was the most over-represented Gene Ontology category for the identified DEPs. Pathway profiling suggested bacterial infections to be linked to aggressive VSCC phenotypes. High Mobility Group AT-Hook 2 (HMGA2) and Proteinase 3 (PRTN3) were revealed as proteins predicting VSCC progression. HMGA2 and PRTN3 abundances are associated with an aggressive phenotype, and hold promise as markers for VSCC patient stratification. It appears that vulvovaginal microflora disturbances trigger an inflammatory response contributing to cancer progression, suggesting that bacterial rather than viral infection status should be considered in the development of targeted therapies in VSCC

Continuous subcutaneous insulin infusion reduces neonatal risk in pregnant women with type 1 diabetes mellitus

Objectives: An attempt was made to demonstrate the superiority of the treatment model using continuous subcutaneous insulin infusion (CSII) over multiple daily injections (MDI) of insulin in achieving a successful pregnancy outcome and good newborn’s condition in patients with type 1 diabetes.  Material and methods: The study included 297 infants born to type 1 diabetic patients; 175 patients were treated with MDI and 122 with CSII.  Maternal metabolic control during pregnancy, gestational weight gain, insulin requirements, pregnancy outcome and neonatal status were compared between MDI and CSII arm.  The composite adverse neonatal outcome was diagnosed if at least one of the following was found: abnormal birth weight (LGA or SGA), congenital malformation, miscarriage, intrauterine fetal death, emergency CS due to fetal risk, iatrogenic prematurity, RDS, hypoglycemia, hyperbilirubinemia, and the postpartum pH in the umbilical artery ≤ 7.1.  Results: The studied groups did not differ regarding gestational week at delivery, a proportion of births at full term, preterm births, miscarriages, or late pregnancy losses (intrauterine fetal death &gt; 22 weeks). Newborns of mothers treated with CSII showed lower incidence of neonatal complications (composite adverse neonatal outcome) compared to those of mothers treated with MDI (60% vs 74%, respectively; p = 0.01). We did not find any association between the mode of treatment and composite adverse maternal outcome.  Conclusions: The use of CSII in the treatment of pregnant women with type 1 diabetes was associated with reduced number of neonatal complications presented as neonatal composite outcome but had no influence on maternal outcome

Barriers to cervical cancer screening exist despite integrating HIV and gynaecological services for HIV-positive women in Poland

Objectives: HIV-positive women are at increased risk of HPV infection and cervical cancer. European and national guidelines advise yearly screening for cervical cancer, however due to the lack of a central registration of HIV infected persons there is a gap in offering such care through general healthcare services in many countries, including Poland. Material and methods: In response to the above limitations, integrated gynaecological care (IGC) was established at the HIV Out-Patient Clinic in Warsaw. We analysed data from January 2007 to May 2014. Logistic regression models were used to identify factors associated with not using IGC by patients. Results: Two hundred and forty women were registered in the observation period:59.6% infected through sexual contact, 18.7% through IDUs, 19.2 % through unknown causes and 2.5% by other (two were vertically infected). The median follow-up time was 2.35 (IQR 0.9–4.5) years and 78.3% were on combination antiretroviral therapy (cART). In total 145 of the women (60.4%) used IGC, from 72.1% of those registered in 2007 to 27.3% registered in 2014. There were in total 1075 gynaeco­logical visits and 254 cervical cytology tests performed. Seventy-five (51.7%) women were tested for HPV infection. Fac­tors decreasing the odds of not using IGC identified by multivariate regression models were being on cART (OR 0.25 [IQR: 0.10–0.59]; p = 0.003) and longer time of observation (0.69 [0.58–0.83]; p = 0.0001). Conclusions: The utilisation of IGC was very high, but with a delay in commencing the IGC. Women on cART and with longer periods of follow-up had lower odds of not using IGC. A screening approach for women not yet on cART, or newly registered in the clinics, needs special attention

Occupational exposure to biological agents in Polish paramedics: a narrative review

INTRODUCTION: Research into occupational exposure to biological pathogens during medical personnel workis to a small degree concerned with paramedics. Coming in contact with biological pathogens, like HIV, HCVand HBV viruses, tubercle bacilli, or recently the SARS-CoV-2 virus in the workplace is a contamination risk.This study aims to analyze the occupational exposure of paramedics to biological pathogens at work, thepossibilities of paramedics developing contagious diseases as occupational illnesses, and the prophylaxisthis involves.MATERIAL AND METHODS: The publication was prepared on the basis of a literature review of works availablein the PubMed, SCOPUS, and Google Scholar databases, and on websites of institutions functioning in thearea of public health.BRIEF DESCRIPTION OF THE STATE OF KNOWLEDGE: In Poland, in 2020, there were 1 255 625 cases of SARSCoV-2 registered, 3020 of which are cases found in paramedics, and 12 524 cases of Lyme borreliosis,990 HBV cases, 942 HCV cases, 934 HIV cases. In 2020, there were 1861 occupational diseases, 504 of whichwere contagious or parasitic. Approximately 37 000 needlesticks are estimated to happen every year in medicalfacilities. 40% to 80% of the people who got injuries or cut in the workplace did not report the incident.CONCLUSIONS: There is a need to implement prophylactic and preventative measures to prevent occupationalneedlestick injuries and blood-borne infections amongst paramedics. Paramedics show insufficientknowledge of their ability to apply for an occupational disease diagnosis caused by exposure to biologicalpathogens present in the work environment

Mutant CAG Repeats Effectively Targeted by RNA Interference in SCA7 Cells

Spinocerebellar ataxia type 7 (SCA7) is a human neurodegenerative polyglutamine (polyQ) disease caused by a CAG repeat expansion in the open reading frame of the ATXN7 gene. The allele-selective silencing of mutant transcripts using a repeat-targeting strategy has previously been used for several polyQ diseases. Herein, we demonstrate that the selective targeting of a repeat tract in a mutant ATXN7 transcript by RNA interference is a feasible approach and results in an efficient decrease of mutant ataxin-7 protein in patient-derived cells. Oligonucleotides (ONs) containing specific base substitutions cause the downregulation of the ATXN7 mutant allele together with the upregulation of its normal allele. The A2 ON shows high allele selectivity at a broad range of concentrations and also restores UCHL1 expression, which is downregulated in SCA7

Inflammatory Proteins HMGA2 and PRTN3 as Drivers of Vulvar Squamous Cell Carcinoma Progression

Current knowledge on the biology of squamous cell vulvar carcinoma (VSCC) is limited. We aimed to identify protein markers of VSCC tumors that would permit to stratify patients by progression risk. Early-stage tumors from patients who progressed (progVSCC) and from those who were disease-free (d-fVSCC) during follow-up, along with normal vulvar tissues were examined by mass spectrometry-based proteomics. Differentially expressed proteins (DEPs) were then verified in solid tissues and blood samples of patients with VSCC tumors and vulvar premalignant lesions. In progVSCC vs. d-fVSCC tumors, the immune response was the most over-represented Gene Ontology category for the identified DEPs. Pathway profiling suggested bacterial infections to be linked to aggressive VSCC phenotypes. High Mobility Group AT-Hook 2 (HMGA2) and Proteinase 3 (PRTN3) were revealed as proteins predicting VSCC progression. HMGA2 and PRTN3 abundances are associated with an aggressive phenotype, and hold promise as markers for VSCC patient stratification. It appears that vulvovaginal microflora disturbances trigger an inflammatory response contributing to cancer progression, suggesting that bacterial rather than viral infection status should be considered in the development of targeted therapies in VSCC