Nephronophthisis-related ciliopathies (NPHP-RCs) are a group of inherited diseases that are associated with defects in primary cilium structure and function. To identify genes mutated in NPHP-RC, we ...performed homozygosity mapping and whole-exome sequencing for >100 individuals, some of whom were single affected individuals born to consanguineous parents and some of whom were siblings of indexes who were also affected by NPHP-RC. We then performed high-throughput exon sequencing in a worldwide cohort of 800 additional families affected by NPHP-RC. We identified two ADAMTS9 mutations (c.4575_4576del p.Gln1525Hisfs∗60 and c.194C>G p.Thr65Arg) that appear to cause NPHP-RC. Although ADAMTS9 is known to be a secreted extracellular metalloproteinase, we found that ADAMTS9 localized near the basal bodies of primary cilia in the cytoplasm. Heterologously expressed wild-type ADAMTS9, in contrast to mutant proteins detected in individuals with NPHP-RC, localized to the vicinity of the basal body. Loss of ADAMTS9 resulted in shortened cilia and defective sonic hedgehog signaling. Knockout of Adamts9 in IMCD3 cells, followed by spheroid induction, resulted in defective lumen formation, which was rescued by an overexpression of wild-type, but not of mutant, ADAMTS9. Knockdown of adamts9 in zebrafish recapitulated NPHP-RC phenotypes, including renal cysts and hydrocephalus. These findings suggest that the identified mutations in ADAMTS9 cause NPHP-RC and that ADAMTS9 is required for the formation and function of primary cilia.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UILJ, UL, UM, UPCLJ, UPUK, ZAGLJ, ZRSKP
Galloway‐Mowat syndrome (GAMOS) is a phenotypically heterogeneous disorder characterized by neurodevelopmental defects combined with renal‐glomerular disease, manifesting with proteinuria. To ...identify additional monogenic disease causes, we here performed whole exome sequencing (WES), linkage analysis, and homozygosity mapping in three affected siblings of an Indian family with GAMOS. Applying established criteria for variant filtering, we identify a novel homozygous splice site mutation in the gene WDR4 as the likely disease‐causing mutation in this family. In line with previous reports, we observe growth deficiency, microcephaly, developmental delay, and intellectual disability as phenotypic features resulting from WDR4 mutations. However, the newly identified allele additionally gives rise to proteinuria and nephrotic syndrome, a phenotype that was never reported in patients with WDR4 mutations. Our data thus expand the phenotypic spectrum of WDR4 mutations by demonstrating that, depending on the specific mutated allele, a renal phenotype may be present. This finding suggests that GAMOS may occupy a phenotypic spectrum with other microcephalic diseases. Furthermore, WDR4 is an additional example of a gene that encodes a tRNA modifying enzyme and gives rise to GAMOS, if mutated. Our findings thereby support the recent observation that, like neurons, podocytes of the renal glomerulus are particularly vulnerable to cellular defects resulting from altered tRNA modifications.
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BFBNIB, FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Abstract
Background
Nephrotic syndrome (NS), a chronic kidney disease, is characterized by significant loss of protein in the urine causing hypoalbuminemia and edema. In general, ∼15% of ...childhood-onset cases do not respond to steroid therapy and are classified as steroid-resistant NS (SRNS). In ∼30% of cases with SRNS, a causative mutation can be detected in one of 44 monogenic SRNS genes. The gene LAMA5 encodes laminin-α5, an essential component of the glomerular basement membrane. Mice with a hypomorphic mutation in the orthologous gene Lama5 develop proteinuria and hematuria.
Methods
To identify additional monogenic causes of NS, we performed whole exome sequencing in 300 families with pediatric NS. In consanguineous families we applied homozygosity mapping to identify genomic candidate loci for the underlying recessive mutation.
Results
In three families, in whom mutations in known NS genes were excluded, but in whom a recessive, monogenic cause of NS was strongly suspected based on pedigree information, we identified homozygous variants of unknown significance (VUS) in the gene LAMA5. While all affected individuals had nonsyndromic NS with an early onset of disease, their clinical outcome and response to immunosuppressive therapy differed notably.
Conclusion
We here identify recessive VUS in the gene LAMA5 in patients with partially treatment-responsive NS. More data will be needed to determine the impact of these VUS in disease management. However, familial occurrence of disease, data from genetic mapping and a mouse model that recapitulates the NS phenotypes suggest that these genetic variants may be inherited factors that contribute to the development of NS in pediatric patients.
Until recently, morpholino oligonucleotides have been widely employed in zebrafish as an acute and efficient loss-of-function assay. However, off-target effects and reproducibility issues when ...compared to stable knockout lines have compromised their further use. Here we employed an acute CRISPR/Cas approach using multiple single guide RNAs targeting simultaneously different positions in two exemplar genes (osgep or tprkb) to increase the likelihood of generating mutations on both alleles in the injected F0 generation and to achieve a similar effect as morpholinos but with the reproducibility of stable lines. This multi single guide RNA approach resulted in median likelihoods for at least one mutation on each allele of >99% and sgRNA specific insertion/deletion profiles as revealed by deep-sequencing. Immunoblot showed a significant reduction for Osgep and Tprkb proteins. For both genes, the acute multi-sgRNA knockout recapitulated the microcephaly phenotype and reduction in survival that we observed previously in stable knockout lines, though milder in the acute multi-sgRNA knockout. Finally, we quantify the degree of mutagenesis by deep sequencing, and provide a mathematical model to quantitate the chance for a biallelic loss-of-function mutation. Our findings can be generalized to acute and stable CRISPR/Cas targeting for any zebrafish gene of interest.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Introduction & Objective: For people with type 2 diabetes (PwT2D), daily insulin regimens can be burdensome, difficult to adhere to, and a key barrier to initiating insulin therapy. Despite ...developments in once-weekly basal insulins, little is known about PwT2D preferences for administration frequency or its importance relative to other insulin features. Methods: An online discrete choice experiment survey was administered to adults with T2D in the United States. Each respondent completed 8 questions offering a choice between experimentally designed pairs of hypothetical, long-acting insulins that varied by 6 attributes: reduction in A1c level after 6 months, daily time in range, number of serious low blood sugar events, number of nighttime low blood sugar events, weight change over 6 months, and administration frequency. A fixed choice question directly elicited preferences for flexible weekly over daily insulin, holding all other attributes equal. Data were analyzed with random-parameter logits. Results: A total of 466 people (mean: age = 57; A1c = 7.5%; 59% female) completed the survey. They were on a basal/bolus (33.3%), basal-only (34.3%), or insulin-naïve (32.4%) regimen. Relative to other attributes, people placed most importance on avoiding a 10-pound weight change, followed by the largest changes in the number of serious and nighttime low blood sugar events and achieving the longest time in range. There was significant heterogeneity in preference: insulin-naïve respondents strongly preferred weekly administration, whether on the same day each week or a flexible weekly schedule, over daily (p < 0.05). In the fixed-choice question, most (67.6%) preferred flexible weekly over daily dosing. Conclusion: Overall, the most important attributes were weight change due to insulin and number of serious hypoglycemic events. PwT2D preferred weekly over daily basal insulin when all other treatment attributes were equal. The dosing flexibility could be crucial for the insulin naïve. Disclosure F. Gelsey: Employee; Eli Lilly and Company. Stock/Shareholder; Eli Lilly and Company. M. Perez-Nieves: Employee; Eli Lilly and Company. Stock/Shareholder; Eli Lilly and Company. C.M. Vass: Other Relationship; Eli Lilly and Company. Consultant; National Institute for Health and Care Excellence (NICE). J. Poon: Employee; Eli Lilly and Company. Stock/Shareholder; Eli Lilly and Company. D. Schapiro: Employee; Eli Lilly and Company. A. Pierce: None. C. Mansfield: None.
Introduction & Objective: Once-weekly insulin has the potential to increase willingness to initiate and adhere to insulin therapies in people with type 2 diabetes requiring basal-only insulin ...therapy. However, the role of once-weekly insulin in people already treated with basal-bolus multiple daily injection regimens, such as people with type 1 diabetes (PwT1D), is less understood. This study aimed to quantify the preferences of PwT1D for novel long-acting insulins. Methods: A discrete choice experiment survey was administered to PwT1D in the United States. The survey included 8 questions offering a choice between experimentally designed pairs of hypothetical, long-acting insulins that varied by 6 attributes: reduction in A1c level after 6 months, daily time in range (TIR), number of serious low blood sugar events, number of nighttime low blood sugar events, weight change over 6 months, and administration frequency. A fixed choice question directly elicited preferences for flexible weekly over daily insulin, holding all other attributes equal. Data were analyzed with random-parameter logits. Results: The survey was completed by 200 PwT1D (mean: age = 41 years; A1c = 7.6%; 57% female). They were on a basal (100%), mealtime bolus (98%), or intermediate or premixed insulin (2%). Across attributes, respondents ranked achieving the longest TIR as the most important change, followed by equal importance on the largest change in the number of serious low blood sugar events per month, avoiding a 10 pound weight change, the largest change in the number of nighttime low blood sugar events per month, and weekly over daily basal administration. The largest reduction in HbA1c was the least important attribute in respondents’ decisions. In the fixed-choice question, 73% of people preferred flexible weekly over daily dosing for basal insulin. Conclusion: The most important attributes were changes in TIR and number of serious hypoglycemic events. PwT1D preferred weekly over daily basal insulin when all other attributes were equal. Disclosure D. Schapiro: Employee; Eli Lilly and Company. M. Perez-Nieves: Employee; Eli Lilly and Company. Stock/Shareholder; Eli Lilly and Company. C.M. Vass: Other Relationship; Eli Lilly and Company. Consultant; National Institute for Health and Care Excellence (NICE). J. Poon: Employee; Eli Lilly and Company. Stock/Shareholder; Eli Lilly and Company. F. Gelsey: Employee; Eli Lilly and Company. Stock/Shareholder; Eli Lilly and Company. A. Pierce: None. C. Mansfield: None.
Background: The impact of BMI on weight (weight) loss in people with T2D differs due to varying modes of weight loss interventions. In this study, we aimed to examine the association of initial BMI ...and identify predictors of weight loss in people with T2D in the US using real-world data. Methods: Adults with T2D in the IQVIA Ambulatory EMR and PharMetrics Plus databases were included in this retrospective study (2010-2019). Index date was date of first observed weight value after T2D diagnosis (index weight). Included individuals had an additional weight value at 1-year (±90 days) post index. People were categorized by their BMI (in kg/m2) at index date (index BMI): 18.5- <25 (normal weight), 25- <30 (overweight), 30- <35 (Class 1 Obesity), 35- <40 (Class 2 Obesity), >40 (Class 3 Obesity). Factors associated with weight loss were evaluated for people who lost >10%-15% or >15% of their index weight at 1-year follow-up using multivariate logistic regression. Results: Of the 1,052,195 people evaluated, 10%, 27%, 29%, 18%, and 16% were categorized into the 5 index BMI categories, respectively. Across the 5 categories, percentage of people with >15% weight loss increased from 0.6% to 4.2%. BMI >40 kg/m2 had the strongest impact on weight loss, among BMI categories (OR=9.1, 95% CI=5.1-16.1). BMI 18.5- <25 kg/m2 did not have a significant impact on weight loss. Overall, the strongest predictor was presence of bariatric surgery (OR=16.3, 95% CI=15.2-17.6), followed by use of antidepressants (OR=1.3, 95% CI=1.2-1.4), being female (OR=1.3, 95% CI=1.2-1.3), use of sulfonylureas (OR=1.2, 95% CI=1.2-1.2), SGLT-2i (OR=1.1, 95% CI=1.1-1.2), and DPP-4i (OR=1.1, 95% CI=1.1-1.2). Similar results were observed for people with >10%-15% weight loss (data not shown). Conclusions: Higher index BMI (>30 kg/m2) was associated with greater weight loss. Prior bariatric surgery, gender, use of antidepressants, sulfonylureas, and DPP-4i were significantly associated with weight loss. The results help in understanding considerations for weight management in people with T2D.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Background: Treatment guidelines recommend >5% sustained weight loss for people with type 2 diabetes (T2D) with overweight/obesity to improve glycemic control and metabolic parameters. We describe ...the impact of weight change on metabolic parameters associated with T2D in the US using real-world data. Methods: A retrospective, longitudinal study was conducted using the IQVIA Ambulatory EMR and the PharMetrics Plus databases (2010-2019). The date of the first observed weight value (i.e., index weight) was defined as the index date. Adults with T2D who had an additional weight value at 1 year (± 90 days) after index date were included. The following metabolic parameters were evaluated at index and 1 year post index: low-density lipoprotein cholesterol (LDL), highdensity lipoprotein cholesterol (HDL), total cholesterol, triglycerides, alanine aminotransferase (ALT), and aspartate aminotransferase (AST). People were grouped based on weight change from index to 1-year follow-up: >3% increase, ± 3% increase/decrease, >3 - <5% decrease, >5 - <10% decrease, >10 - 15% decrease, and >15% decrease. Data are presented as percentages (categorical) and mean change from index date (continuous). Results: Among the included population (N=1,061,354, mean index weight= 95.9 kg), 20%, 54%, 11%, 11%, 3%, and 2% were in the respective weight change groups described above. The largest mean change from index date in LDL (-10.2 mg/dL) and total cholesterol (-15.1 mg/dL), was observed in people with >10 - <15% weight decrease. Greater reduction was observed in the other metabolic parameters, as weight change progressed from >3% increase to >15% decrease: HDL (range: -0.2 to 5.1 mg/dL), triglycerides (range: -5.9 to -58.4 mg/dL), ALT (range: 0.7 to -9.9 units/liter) and AST (range: 0.4 to -5.2 units/liter). Conclusions: Our findings suggest that greater improvements in metabolic parameters may be associated with higher weight reduction after 1 year among adults with T2D.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
Background: In the past 5 years, type 2 diabetes practice guidelines have shifted to prioritize agents with cardiovascular (CV) and renal benefit. This study assessed whether real-world medication ...use has changed in line with these evolving guidelines.
Methods: We retrospectively examined the treatment patterns (12 months pre and post) of people with T2D (PwT2D) who initiated basal insulin (BI) in either 2015 (cohort 1) , 2017 (cohort 2) , or 2019 (cohort 3) using IBM® MarketScan® Databases. Between-group differences were analyzed via t-test or chi-square as appropriate.
Results: In the study period, 6,396,441 PwT2D were identified. The number of PwT2D initiating BI was 17,801/948,805 (1.88%) in 2015 vs. 10,870/622,455 (1.75%) in 2019 and a progressively larger share of BI initiators had prior GLP-1RA use (14.8% in cohort 1 vs. 25.2% in cohort 3; p < 0.001) or SGLT-2i use (11.4% in cohort 1 vs. 20.5% in cohort 3; p < 0.001) . Furthermore, in patients without prior GLP-1RA, SGLT-2i, or bolus use who initiated GLP-1RA or SGLT-2i in the first year of BI, time to first GLP-1RA (132.4 days in cohort 1 vs. 120.5 days in cohort 3; p = 0.02) or SGLT-2i (131.5 days in cohort 1 vs. 113.3 days in cohort 3; p = 0.002) decreased.
Conclusions: Real-world data suggests growing and earlier use of GLP-1RA and SGLT-2i among BI users, and a growing proportion of PwT2D on BI-GLP-1RA combination treatment, in line with changing practice guidelines.
Disclosure
D. Schapiro: Employee; Eli Lilly and Company. A. Meeks: Employee; Eli Lilly and Company. D. Liu: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. F. Gelsey: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. R. Juneja: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. M. Perez-nieves: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. A. Huang: None.
Funding
Eli Lilly and Company
Limited US real-world evidence exists on the effect of substantial changes in weight on HbA1c among people with T2D. People with T2D were identified from 2010-2018 in the IQVIA Ambulatory EMR ...database. The date of the first observed weight value was defined as the index date and patients were required to have an additional weight value at 1 year (±90 days) post-index date. Patients were stratified by weight change between index and 1 year post-index date. HbA1c was evaluated at index, 6 months, and 1 year post-index date. Of the 1,061,354 patients evaluated, 20.2% gained weight, 53.9% had no weight change, and 10.6% lost 3-5%, 10.7% lost 5-10%, 2.8% lost 10-15%, and 1.7% lost ≥15% of their weight at the index date (i.e., index weight) at 1 year of follow-up. Of note, patients with the largest weight loss had the highest index weight (range: 203.7lb in patients with weight gain to 245.9lb in patients with ≥15% weight loss) . Mean index HbA1c was similar across subgroups as was the percent of patients with index HbA1c <7.0%. At both 6 months and 1 year post-index date, patients with ≥15% weight loss had the largest reduction in HbA1c and the highest percent of patients with HbA1c <7.0%.
In summary, HbA1c reduction was greater with increasing amount of weight lost during the one year of follow-up. Patients with weight loss ≥15% reduced HbA1c by an average of 1.2% of the index value at 1 year of follow-up.
Disclosure
S.Shinde: None. V.Thieu: None. A.Kwan: Employee; Eli Lilly and Company, Stock/Shareholder; Eli Lilly and Company. K.F.Houghton: None. D.Schapiro: Employee; Eli Lilly and Company. J.Meyers: Other Relationship; Eli Lilly and Company.
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