Association of Candidate Gene Polymorphisms With Chronic Kidney Disease: Results of a Case-Control Analysis in the Nefrona Cohort

Chronic kidney disease (CKD) is a major risk factor for end-stage renal disease, cardiovascular disease and premature death. Despite classical clinical risk factors for CKD and some genetic risk factors have been identified, the residual risk observed in prediction models is still high. Therefore, new risk factors need to be identified in order to better predict the risk of CKD in the population. Here, we analyzed the genetic association of 79 SNPs of proteins associated with mineral metabolism disturbances with CKD in a cohort that includes 2, 445 CKD cases and 559 controls. Genotyping was performed with matrix assisted laser desorption ionizationtime of flight mass spectrometry. We used logistic regression models considering different genetic inheritance models to assess the association of the SNPs with the prevalence of CKD, adjusting for known risk factors. Eight SNPs (rs1126616, rs35068180, rs2238135, rs1800247, rs385564, rs4236, rs2248359, and rs1564858) were associated with CKD even after adjusting by sex, age and race. A model containing five of these SNPs (rs1126616, rs35068180, rs1800247, rs4236, and rs2248359), diabetes and hypertension showed better performance than models considering only clinical risk factors, significantly increasing the area under the curve of the model without polymorphisms. Furthermore, one of the SNPs (the rs2248359) showed an interaction with hypertension, being the risk genotype affecting only hypertensive patients. We conclude that 5 SNPs related to proteins implicated in mineral metabolism disturbances (Osteopontin, osteocalcin, matrix gla protein, matrix metalloprotease 3 and 24 hydroxylase) are associated to an increased risk of suffering CKD

Familial hypercholesterolaemia in children and adolescents from 48 countries: a cross-sectional study

Background: Approximately 450 000 children are born with familial hypercholesterolaemia worldwide every year, yet only 2·1% of adults with familial hypercholesterolaemia were diagnosed before age 18 years via current diagnostic approaches, which are derived from observations in adults. We aimed to characterise children and adolescents with heterozygous familial hypercholesterolaemia (HeFH) and understand current approaches to the identification and management of familial hypercholesterolaemia to inform future public health strategies. Methods: For this cross-sectional study, we assessed children and adolescents younger than 18 years with a clinical or genetic diagnosis of HeFH at the time of entry into the Familial Hypercholesterolaemia Studies Collaboration (FHSC) registry between Oct 1, 2015, and Jan 31, 2021. Data in the registry were collected from 55 regional or national registries in 48 countries. Diagnoses relying on self-reported history of familial hypercholesterolaemia and suspected secondary hypercholesterolaemia were excluded from the registry; people with untreated LDL cholesterol (LDL-C) of at least 13·0 mmol/L were excluded from this study. Data were assessed overall and by WHO region, World Bank country income status, age, diagnostic criteria, and index-case status. The main outcome of this study was to assess current identification and management of children and adolescents with familial hypercholesterolaemia. Findings: Of 63 093 individuals in the FHSC registry, 11 848 (18·8%) were children or adolescents younger than 18 years with HeFH and were included in this study; 5756 (50·2%) of 11 476 included individuals were female and 5720 (49·8%) were male. Sex data were missing for 372 (3·1%) of 11 848 individuals. Median age at registry entry was 9·6 years (IQR 5·8-13·2). 10 099 (89·9%) of 11 235 included individuals had a final genetically confirmed diagnosis of familial hypercholesterolaemia and 1136 (10·1%) had a clinical diagnosis. Genetically confirmed diagnosis data or clinical diagnosis data were missing for 613 (5·2%) of 11 848 individuals. Genetic diagnosis was more common in children and adolescents from high-income countries (9427 [92·4%] of 10 202) than in children and adolescents from non-high-income countries (199 [48·0%] of 415). 3414 (31·6%) of 10 804 children or adolescents were index cases. Familial-hypercholesterolaemia-related physical signs, cardiovascular risk factors, and cardiovascular disease were uncommon, but were more common in non-high-income countries. 7557 (72·4%) of 10 428 included children or adolescents were not taking lipid-lowering medication (LLM) and had a median LDL-C of 5·00 mmol/L (IQR 4·05-6·08). Compared with genetic diagnosis, the use of unadapted clinical criteria intended for use in adults and reliant on more extreme phenotypes could result in 50-75% of children and adolescents with familial hypercholesterolaemia not being identified. Interpretation: Clinical characteristics observed in adults with familial hypercholesterolaemia are uncommon in children and adolescents with familial hypercholesterolaemia, hence detection in this age group relies on measurement of LDL-C and genetic confirmation. Where genetic testing is unavailable, increased availability and use of LDL-C measurements in the first few years of life could help reduce the current gap between prevalence and detection, enabling increased use of combination LLM to reach recommended LDL-C targets early in life

Trichrome-stained sections of the vaginal wall of <i>Fbln5</i>^<i>-/-</i> mice treated with HG (A) or HG+fibulin-5 (10 μg/ml).

<p>Note increased thickness of epithelial and stromal layers in animals treated with fibulin-5. Bar = 20 μm.</p

Thermosensitive hydrogels deliver bioactive protein to the vaginal wall - Fig 4

<p><b>Effect of HG ± fibulin-5 on MMP-9 activity in vaginal stromal cells from WT (A) or Fbln5</b>^<b>-/-</b><b>(B) mice. HG,</b> hydrogel; <b>Fib-5</b>_<b>5,</b> recombinant fibulin-5 5 μg/ml; <b>Fib-5</b>_<b>10,</b> recombinant fibulin-5 10 μg/ml. <b>-Ctl,</b> vaginal tissues from MMP-9 KO mice; <b>+Ctl,</b> vaginal tissues from Fbln5^-/- mice.</p

Long-term effects of HG ± fibulin-5 on vaginal MMP-9 in <i>Fbln5</i> KO mice treated for 14–28 d.

<p><b>A.</b> Cumulative results of MMP-9 activity in vaginal tissues from 38 mice treated with HG ± fibulin-5 (10 μg/ml) for 2–4 weeks. *P < 0.05 compared with HG alone, ANOVA. <b>B.</b> Immunoblot analysis of urea-extracted protein from vaginal tissues of Fbln5^-/- mice treated with HG or HG + FBLN5 (10 μg/ml) x 2 weeks. The left blot was incubated with <b>anti- His Tag</b> antibody whereas the right blot was incubated with anti-fibulin-5. Positive controls on each blot are recombinant fibulin-5 (rFBLN5, 200 ng). <b>Bl</b>, blank.</p

Thermosensitive property of the polymer solution.

<p><b>A</b>. Transmittance of the polymer aqueous solution (0.5 wt%) as a function of temperature. <b>B</b>. Thermally-induced gelation of the HG aqueous solution (25 wt%).</p

Effect of HG ± fibulin-5 on vaginal MMP-9 and MMP-2 in <i>Fbln5</i> KO mice treated for 7 d.

<p><b>A.</b> Gelatin zymography with protein extracts (10 μg/lane) from adult <i>Fbln5</i>^<i>-/-</i> mice injected with hydrogel alone (HG), HG incorporated with fibulin-5 (5 ug/ml), or fibulin-5 alone x 7 d. <i>Mmp9</i> KO was used as a neg ctl. <b>B.</b> Cumulative results of 34 mice treated with HG ± various doses of fibulin-5 x 7 d. *P < 0.05 compared with HG alone, ANOVA.</p

<i>In vitro</i> cell compatibility and <i>in vivo</i> tissue compatibility of HG was assessed.

<p>(A) HG was incubated with DMEM media for different periods of time (1, 3, 5 and 7 days) to generate conditioned media. The cell compatibility of the conditioned media was then evaluated using 3T3 fibroblasts and MTT assay. (B) Balb/c mice were subcutaneously implanted with HG or saline as the control. After implantation for 7 days, the animals were sacrificed and the implant-surrounding tissues were recovered for histological analyses. H&E stain and implant-associated cell numbers support that HG implant exerted minimal tissue response similar to saline control.</p