CFTR interactome mapping using the mammalian membrane two-hybrid high-throughput screening system

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is a chloride and bicarbonate channel in secretory epithelia with a critical role in maintaining fluid homeostasis. Mutations in CFTR are associated with Cystic Fibrosis (CF), the most common lethal autosomal recessive disorder in Caucasians. While remarkable treatment advances have been made recently in the form of modulator drugs directly rescuing CFTR dysfunction, there is still considerable scope for improvement of therapeutic effectiveness. Here, we report the application of a high-throughput screening variant of the Mammalian Membrane Two-Hybrid (MaMTH-HTS) to map the protein-protein interactions of wild-type (wt) and mutant CFTR (F508del), in an effort to better understand CF cellular effects and identify new drug targets for patient-specific treatments. Combined with functional validation in multiple disease models, we have uncovered candidate proteins with potential roles in CFTR function/CF pathophysiology, including Fibrinogen Like 2 (FGL2), which we demonstrate in patient-derived intestinal organoids has a significant effect on CFTR functional expression

Canagliflozin and renal outcomes in type 2 diabetes and nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to <90 ml per minute per 1.73 m2 of body-surface area and albuminuria (ratio of albumin [mg] to creatinine [g], >300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of <15 ml per minute per 1.73 m2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P<0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P<0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Canagliflozin and Renal Outcomes in Type 2 Diabetes and Nephropathy

BACKGROUND Type 2 diabetes mellitus is the leading cause of kidney failure worldwide, but few effective long-term treatments are available. In cardiovascular trials of inhibitors of sodium–glucose cotransporter 2 (SGLT2), exploratory results have suggested that such drugs may improve renal outcomes in patients with type 2 diabetes. METHODS In this double-blind, randomized trial, we assigned patients with type 2 diabetes and albuminuric chronic kidney disease to receive canagliflozin, an oral SGLT2 inhibitor, at a dose of 100 mg daily or placebo. All the patients had an estimated glomerular filtration rate (GFR) of 30 to 300 to 5000) and were treated with renin–angiotensin system blockade. The primary outcome was a composite of end-stage kidney disease (dialysis, transplantation, or a sustained estimated GFR of <15 ml per minute per 1.73 m 2), a doubling of the serum creatinine level, or death from renal or cardiovascular causes. Prespecified secondary outcomes were tested hierarchically. RESULTS The trial was stopped early after a planned interim analysis on the recommendation of the data and safety monitoring committee. At that time, 4401 patients had undergone randomization, with a median follow-up of 2.62 years. The relative risk of the primary outcome was 30% lower in the canagliflozin group than in the placebo group, with event rates of 43.2 and 61.2 per 1000 patient-years, respectively (hazard ratio, 0.70; 95% confidence interval [CI], 0.59 to 0.82; P=0.00001). The relative risk of the renal-specific composite of end-stage kidney disease, a doubling of the creatinine level, or death from renal causes was lower by 34% (hazard ratio, 0.66; 95% CI, 0.53 to 0.81; P<0.001), and the relative risk of end-stage kidney disease was lower by 32% (hazard ratio, 0.68; 95% CI, 0.54 to 0.86; P=0.002). The canagliflozin group also had a lower risk of cardiovascular death, myocardial infarction, or stroke (hazard ratio, 0.80; 95% CI, 0.67 to 0.95; P=0.01) and hospitalization for heart failure (hazard ratio, 0.61; 95% CI, 0.47 to 0.80; P<0.001). There were no significant differences in rates of amputation or fracture. CONCLUSIONS In patients with type 2 diabetes and kidney disease, the risk of kidney failure and cardiovascular events was lower in the canagliflozin group than in the placebo group at a median follow-up of 2.62 years

Characterization of TMEM14 Proteins in the Context of Zebrafish PCP AND Ciliogenesis

The planar cell polarity (PCP) pathway regulates key developmental processes which include convergence and extension (C) movements and the planar organization of motile cilia (1–3). Central members of the PCP pathway include transmembrane proteins such as Van Gogh-like (Vangl). However, despite a large body of research characterizing the PCP pathway, the intracellular trafficking of each factor is poorly understood. This knowledge gap has led to a search for additional factors that act in the regulation of PCP components. In this work, I expand on the characterization of the trans-Golgi network (TGN) localized protein, Tmem14A, an unpublished novel interactor of Vangl1/2 in the context zebrafish PCP and ciliogenesis. Curiously, I found that the loss of Tmem14A in both zygotic and maternal zygotic (MZ) tmem14A mutants did not severely impact the morphogenesis of zebrafish embryos. However, in my analysis of MZtmem14A floor plate (FP) neuroepithelia, I show the planar polarity of motile cilia to be disrupted and the localization of Vangl2 to the basolateral membranes of FP cells to be reduced. This analysis supports the hypothesis that Tmem14A can regulate PCP, possibly through the intracellular transport of Vangl2 from the TGN. The absence of severe developmental defects in MZtmem14A mutants suggests that Tmem14A function may be dispensable in the context of embryonic development. Interestingly, a recent study has provided evidence to suggest that the loss of gene function can be rescued by the upregulation of a functionally related gene, induced by a novel non-sense mediated decay (NMD)-dependent compensatory mechanism (4,5). Intriguingly, I show that MZtmem14A mutants have a near 2-fold upregulation of tmem14cB, an uncharacterized zebrafish-specific Tmem14 family member, suggesting that loss of Tmem14A may be rescued by Tmem14cB. Interestingly, contrary to my initial hypothesis of Tmem14cB alleviating the loss of Tmem14A, MZtmem14A;MZtmem14cB mutants did not develop morphological defects. Remarkably, the loss of Tmem14cB rescued the translational polarity and basolateral Vangl2 defects of FP cells in MZtmem14A mutants. These observations suggest that the upregulation of tmem14cB in MZtmem14A mutants has a deleterious effect on FP planar polarity and implicates Tmem14cB as a novel regulator of Vangl2 and zebrafish planar polarity.Ph.D

Distancing from a stigmatized identity: Explaining hostility by marginalized racial groups toward new immigrants

Discrimination is often perceived as stemming from outgroups. The UN Sustainable Development Goal 10 focused on reducing inequalities calls attention also to intragroup hostilities. In the US, intragroup hostilities between Latinos/as might occur if disassociation from a stigmatized sub-group protects one’s status. This chapter tests potential disassociation effects by examining whether US Latinos/as distance themselves from an associated stigmatized identity by supporting adverse policies regarding Latino/a immigrants. Two studies (n=273 and n=8634) found that citizenship status was linked to support for adverse policies: more US-born Latinos/as considered immigrants a burden than Latinos/as of unknown status or non-citizens. Some Latino/a citizens might cut off reflected failure associated with being an immigrant because distancing might support coping with cultural demands of US residence and distancing from recent immigrants might prevent transference of negative stereotypes. As inequalities increase overall in the post-COVID-19 era, intragroup bias may worsen outcomes for stigmatized sub-groups

Evaluation of Measures of Upper Limb Functioning and Disability in People With Parkinson Disease: A Systematic Review

To identify measurement tools used for upper limb evaluation in people with Parkinson disease ( PD ), to summarize the content of each tool using the International Classification of Functioning, Disability and Health, and to examine the reliability, validity, clinical utility, and responsiveness of the measurement tools specific to this clinical group