siRNA Delivery to the Glomerular Mesangium Using Polycationic Cyclodextrin Nanoparticles Containing siRNA

There is an urgent need for new therapies that can halt or reverse the course of chronic kidney disease with minimal side-effect burden on the patient. Small interfering RNA (siRNA) nanoparticles are new therapeutic entities in clinical development that could be useful for chronic kidney disease treatment because they combine the tissue-specific targeting properties of nanoparticles with the gene-specific silencing effects of siRNA. Recent reports have emerged demonstrating that the kidney, specifically the glomerulus, is a readily accessible site for nanoparticle targeting. Here, we explore the hypothesis that intravenously administered polycationic cyclodextrin nanoparticles containing siRNA (siRNA/CDP-NPs) can be used for delivery of siRNA to the glomerular mesangium. We demonstrate that siRNA/CDP-NPs localize to the glomerular mesangium with limited deposition in other areas of the kidney after intravenous injection. Additionally, we report that both mouse and human mesangial cells rapidly internalize siRNA/CDP-NPs in vitro and that nanoparticle uptake can be enhanced by attaching the targeting ligands mannose or transferrin to the nanoparticle surface. Lastly, we show knockdown of mesangial enhanced green fluorescent protein expression in a reporter mouse strain following iv treatment with siRNA/CDP-NPs. Altogether, these data demonstrate the feasibility of mesangial targeting using intravenously administered siRNA/CDP-NPs

Far-infrared and sub-millimetre imaging of HD 76582's circumstellar disc

Debris discs, the tenuous rocky and icy remnants of planet formation, are believed to be evidence for planetary systems around other stars. The JCMT/SCUBA-2 debris disc legacy survey ‘SCUBA-2 Observations of Nearby Stars’ (SONS) observed 100 nearby stars, amongst them HD 76582, for evidence of such material. Here, we present imaging observations by JCMT/SCUBA-2 and Herschel/PACS at sub-millimetre and far-infrared wavelengths, respectively. We simultaneously model the ensemble of photometric and imaging data, spanning optical to sub-millimetre wavelengths, in a self-consistent manner. At far-infrared wavelengths, we find extended emission from the circumstellar disc providing a strong constraint on the dust spatial location in the outer system, although the angular resolution is too poor to constrain the interior of the system. In the sub-millimetre, photometry at 450 and 850 µm reveals a steep fall-off that we interpret as a disc dominated by moderately sized dust grains (amin = 36 µm), perhaps indicative of a non-steady-state collisional cascade within the disc. A disc architecture of three distinct annuli, comprising an unresolved component at 20 au and outer components at 80 and 270 au, along with a very steep particle size distribution (γ = 5), is proposed to match the observations

Bayesian analysis to identify new star candidates in nearby young stellar kinematic groups

We present a new method based on a Bayesian analysis to identify new members of nearby young kinematic groups. The analysis minimally takes into account the position, proper motion, magnitude and color of a star, but other observables can be readily added (e.g. radial velocity, distance). We use this method to find new young low-mass stars in the \beta Pictoris (\beta PMG) and AB Doradus (ABDMG) moving groups and in the TW Hydrae (TWA), Tucana-Horologium (THA), Columba, Carina and Argus associations. Starting from a sample of 758 mid-KM (K5V-M5V) stars showing youth indicators such as H\alpha\ and X-ray emission, our analysis yields 215 new highly probable low-mass members of the kinematic groups analyzed. One is in TWA, 37 in \beta PMG, 17 in THA, 20 in Columba, 6 in Carina, 50 in Argus, 33 in ABDMG, and the remaining 51 candidates are likely young but have an ambiguous membership to more than one association. The false alarm rate for new candidates is estimated to be 5% for \beta PMG and TWA, 10% for THA, Columba, Carina and Argus, and 14% for ABDMG. Our analysis confirms the membership of 58 stars proposed in the literature. Firm membership confirmation of our new candidates will require measurement of their radial velocity (predicted by our analysis), parallax and lithium 6708 {\AA} equivalent width. We have initiated these follow-up observations for a number of candidates and we have identified two stars (2MASSJ0111+1526, 2MASSJ0524-1601) as very strong candidate members of the \beta PMG and one strong candidate member (2MASSJ0533-5117) of the THA; these three stars have radial velocity measurements confirming their membership and lithium detections consistent with young age. Finally, we proposed that six stars should be considered as new bona fide members of \beta PMG and ABDMG, one of which being first identified in this work, the others being known candidates from the literature.Comment: Accepted for publication in Ap

Store-Operated Ca^(2+) Channels in Mesangial Cells Inhibit Matrix Protein Expression

Accumulation of extracellular matrix derived from glomerular mesangial cells is an early feature of diabetic nephropathy. Ca^(2+) signals mediated by store–operated Ca^(2+) channels regulate protein production in a variety of cell types. The aim of this study was to determine the effect of store–operated Ca^(2+) channels in mesangial cells on extracellular matrix protein expression. In cultured human mesangial cells, activation of store–operated Ca^(2+) channels by thapsigargin significantly decreased fibronectin protein expression and collagen IV mRNA expression in a dose-dependent manner. Conversely, inhibition of the channels by 2-aminoethyl diphenylborinate significantly increased the expression of fibronectin and collagen IV. Similarly, overexpression of stromal interacting molecule 1 reduced, but knockdown of calcium release–activated calcium channel protein 1 (Orai1) increased fibronectin protein expression. Furthermore, 2-aminoethyl diphenylborinate significantly augmented angiotensin II–induced fibronectin protein expression, whereas thapsigargin abrogated high glucose– and TGF-β1–stimulated matrix protein expression. In vivo knockdown of Orai1 in mesangial cells of mice using a targeted nanoparticle siRNA delivery system resulted in increased expression of glomerular fibronectin and collagen IV, and mice showed significant mesangial expansion compared with controls. Similarly, in vivo knockdown of stromal interacting molecule 1 in mesangial cells by recombinant adeno–associated virus–encoded shRNA markedly increased collagen IV protein expression in renal cortex and caused mesangial expansion in rats. These results suggest that store–operated Ca^(2+) channels in mesangial cells negatively regulate extracellular matrix protein expression in the kidney, which may serve as an endogenous renoprotective mechanism in diabetes

CRLX101 nanoparticles localize in human tumors and not in adjacent, nonneoplastic tissue after intravenous dosing

Nanoparticle-based therapeutics are being used to treat patients with solid tumors. Whereas nanoparticles have been shown to preferentially accumulate in solid tumors of animal models, there is little evidence to prove that intact nanoparticles localize to solid tumors of humans when systemically administered. Here, tumor and adjacent, nonneoplastic tissue biopsies are obtained through endoscopic capture from patients with gastric, gastroesophageal, or esophageal cancer who are administered the nanoparticle CRLX101. Both the pre- and postdosing tissue samples adjacent to tumors show no definitive evidence of either the nanoparticle or its drug payload (camptothecin, CPT) contained within the nanoparticle. Similar results are obtained from the predosing tumor samples. However, in nine of nine patients that were evaluated, CPT is detected in the tumor tissue collected 24–48 h after CRLX101 administration. For five of these patients, evidence of the intact deposition of CRLX101 nanoparticles in the tumor tissue is obtained. Indications of CPT pharmacodynamics from tumor biomarkers such as carbonic anhydrase IX and topoisomerase I by immunohistochemistry show clear evidence of biological activity from the delivered CPT in the posttreatment tumors

Correlating animal and human phase Ia/Ib clinical data with CALAA-01, a targeted, polymer-based nanoparticle containing siRNA

Nanoparticle-based experimental therapeutics are currently being investigated in numerous human clinical trials. CALAA-01 is a targeted, polymer-based nanoparticle containing small interfering RNA (siRNA) and, to our knowledge, was the first RNA interference (RNAi)–based, experimental therapeutic to be administered to cancer patients. Here, we report the results from the initial phase I clinical trial where 24 patients with different cancers were treated with CALAA-01 and compare those results to data obtained from multispecies animal studies to provide a detailed example of translating this class of nanoparticles from animals to humans. The pharmacokinetics of CALAA-01 in mice, rats, monkeys, and humans show fast elimination and reveal that the maximum concentration obtained in the blood after i.v. administration correlates with body weight across all species. The safety profile of CALAA-01 in animals is similarly obtained in humans except that animal kidney toxicities are not observed in humans; this could be due to the use of a predosing hydration protocol used in the clinic. Taken in total, the animal models do appear to predict the behavior of CALAA-01 in humans