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Increased expression of the ATP-binding cassette family member A1 (ABCA1) protein elicits numerous therapeutic effects against type 2 diabetes, Alzheimer’s disease, and other related conditions in preclinical models. However, side effects such as increased lipogenesis and neutropenia have precluded translation of ABCA1-inducing compounds to the clinic. This project utilized a unique phenotypic strategy to develop selective, non-lipogenic ABCA1 inducers as multifunctional therapeutic candidates for Alzheimer’s disease and type 2 diabetes. The approach consisted of chemical synthesis to optimize a screening hit into lead candidates with enhanced potency and efficacy, followed by testing in an array of cell and animal models. Hit-to-lead optimization of the initial screening hit F420 yielded two key lead compounds, CL2-57 and CL3-3. Both lead compounds selectively enhanced expression of ABCA1 over genes promoting hepatic lipogenesis. This selectivity stemmed from combined agonist and antagonist effects at multiple nuclear receptor isoforms, which represented a novel mechanism of action. Additionally, compound CL3-3 displayed a stronger combination of potency for ABCA1 induction and selectivity against lipogenic gene induction than the best previously reported agonists. These two non-lipogenic ABCA1 inducers also enhanced cholesterol efflux and blunted pro-inflammatory signaling in cell-based assays. After in vitro testing, the lead compounds were evaluated for safety and efficacy in mouse models. Both ABCA1 inducers maintained functional selectivity in vivo, enhancing ABCA1 expression and modifying disease phenotypes while avoiding unwanted effects, such as increased triglycerides. Major efficacy readouts included phenotypic, biochemical, and metabolomic analysis of a high-fat diet mouse model. Oral treatment with CL2-57 enhanced glucose homeostasis, reduced adiposity, attenuated inflammation, and reversed metabolic perturbations in mice fed the high-fat diet. Oral treatment with CL3-3 also induced cholesterol transport genes in the brain and periphery, while triglyceride increases were avoided. Testing for effects of CL3-3 on Alzheimer’s-related pathology in additional mouse models is ongoing, but these compounds are promising leads for further development as Alzheimer’s disease and type 2 diabetes therapeutics.

Stochastic fluctuations of the neutron population within a nuclear reactor are typically prevented by operating the core at a sufficient power, since a deterministic behavior of the neutron population is required by automatic safety systems to detect unwanted power excursions. Recent works however pointed out that, under specific circumstances, non-Poissonian patterns could affect neutron spatial distributions. This motivated an international program to experimentally detect and characterize such fluctuations and correlations, which took place in 2017 at the Rensselaer Polytechnic Institute Reactor Critical Facility. The main findings of this program will indeed unveil patchiness in snapshots of neutron spatial distributions -- obtained with a dedicated numerical twin of the reactor -- that support this first experimental characterization of the 'neutron clustering' phenomenon, while a stochastic model based on reaction-diffusion processes and branching random walks will reveal the key role played by the reactor intrinsic sources in understanding neutron spatial correlations., 43 pages, 6 figures

Introduction: Reduced expression of cholesterol transporter ABCA1 is critical in pathogenesis of type 2 diabetes (T2D) and related conditions, such as cardiovascular disease (CVD) and Alzheimer’s disease (AD). Thus, increasing ABCA1 represents a novel therapeutic strategy for these conditions. However, prior drug development efforts have achieved limited success at increasing ABCA1 (controlled by liver X receptor [LXR] β) while avoiding unwanted liver triglyceride production (through LXRα via transcription factor SREBP1c). Hypothesis: We hypothesized that phenotypic screening for selective ABCA1 inducers followed by medicinal chemistry optimization would bypass the isoform selectivity issues encountered in traditional target-based drug discovery and enable development of lead therapeutic candidates with preclinical efficacy and safety. Methods/Results: We screened 20k compounds for ABCA1 and SREBP1c-linked luciferase activity, followed by qPCR to validate and prioritize selective ABCA1-inducing hits. We synthesized ~70 structural analogs of the best hit, achieving substantial EC 50 (4.5 μM to 270 nM) and E max (3.5-fold to 6.0-fold vs. vehicle) improvements in ABCA1 luciferase assay while maintaining selectivity against SREBP1c. Direct binding assays confirmed selectivity for LXRβ vs. LXRα, corroborating cell-based data. Lead compounds enhanced cellular cholesterol efflux, reduced inflammation in vitro , and attenuated high-fat diet (HFD) induced weight gain, insulin resistance, and inflammation in mice. Metabolomics analysis revealed that our lead compound corrected HFD-induced perturbations in liver glucose and fatty acid synthesis. Finally, side effects associated with published LXR agonists - liver steatosis and neutropenia - were not observed with our compound. Conclusions: We established a platform to develop selective ABCA1 inducers as drug candidates. Via this platform, we identified a safe and efficacious lead compound for T2D. Our study also represents the first report of an LXR agonist characterized by metabolomics - a powerful tool to complement biochemical readouts. Continued optimization to improve pharmacokinetic parameters, plus evaluation in CVD and AD models, is ongoing.

[Image: see text] Selective liver X receptor (LXR) agonists have been extensively pursued as therapeutics for Alzheimer’s disease and related dementia (ADRD) and, for comorbidities such as type 2 diabetes (T2D) and cerebrovascular disease (CVD), disorders with underlying impaired insulin signaling, glucose metabolism, and cholesterol mobilization. The failure of the LXR-focused approach led us to pursue a novel strategy to discover nonlipogenic ATP-binding cassette transporter A1 (ABCA1) inducers (NLAIs): screening for ABCA1-luciferase activation in astrocytoma cells and counterscreening against lipogenic gene upregulation in hepatocarcinoma cells. Beneficial effects of LXRβ agonists mediated by ABCA1 include the following: control of cholesterol and phospholipid efflux to lipid-poor apolipoproteins forming beneficial peripheral HDL and HDL-like particles in the brain and attenuation of inflammation. While rare, ABCA1 variants reduce plasma HDL and correlate with an increased risk of ADRD and CVD. In secondary assays, NLAI hits enhanced cholesterol mobilization and positively impacted in vitro biomarkers associated with insulin signaling, inflammatory response, and biogenic properties. In vivo target engagement was demonstrated after oral administration of NLAIs in (i) mice fed a high-fat diet, a model for obesity-linked T2D, (ii) mice administered LPS, and (iii) mice with accelerated oxidative stress. The lack of adverse effects on lipogenesis and positive effects on multiple biomarkers associated with T2D and ADRD supports this novel phenotypic approach to NLAIs as a platform for T2D and ADRD drug discovery.

Background Therapeutic agents with novel mechanisms of action are needed to combat the growing epidemic of type 2 diabetes (T2D) and related metabolic syndromes. Liver X receptor (LXR) agonists possess preclinical efficacy yet produce side effects due to excessive lipogenesis. Anticipating that many beneficial and detrimental effects of LXR agonists are mediated by ABCA1 and SREPB1c expression, respectively, we hypothesized that a phenotypic optimization strategy prioritizing selective ABCA1 induction would identify an efficacious lead compound with an improved side effect profile over existing LXRβ agonists. Methods We synthesized and characterized a novel small molecule for selective induction of ABCA1 vs. SREBP1c in vitro. This compound was evaluated in both wild-type mice and a high-fat diet (HFD) mouse model of obesity-driven diabetes through functional, biochemical, and metabolomic analysis. Findings Six weeks of oral administration of our lead compound attenuated weight gain, glucose intolerance, insulin signaling deficits, and adiposity. Global metabolomics revealed suppression of gluconeogenesis, free fatty acids, and pro-inflammatory metabolites. Target identification linked these beneficial effects to selective LXRβ agonism and PPAR/RXR antagonism. Interpretation Our observations in the HFD model, combined with the absence of lipogenesis and neutropenia in WT mice, support this novel approach to therapeutic development for T2D and related conditions.

The focus on amyloid plaques and neurofibrillary tangles has yielded no Alzheimer’s disease (AD) modifying treatments in the past several decades, despite successful studies in preclinical mouse models. This inconsistency has caused a renewed focus on improving the fidelity and reliability of AD mouse models, with disparate views on how this improvement can be accomplished. However, the interactive effects of the universal biological variables of AD, which include age, APOE genotype, and sex, are often overlooked. Age is the greatest risk factor for AD, while the e4 allele of the human APOE gene, encoding apolipoprotein E, is the greatest genetic risk factor. Sex is the final universal biological variable of AD, as females develop AD at almost twice the rate of males and, importantly, female sex exacerbates the effects of APOE4 on AD risk and rate of cognitive decline. Therefore, this review evaluates the importance of context for understanding the role of APOE in preclinical mouse models. Specifically, we detail how human AD pathology is mirrored in current transgenic mouse models (“What”) and describe the critical need for introducing human APOE into these mouse models (“Who”). We next outline different methods for introducing human APOE into mice (“How”) and highlight efforts to develop temporally defined and location-specific human apoE expression models (“When” and “Where”). We conclude with the importance of choosing the human APOE mouse model relevant to the question being addressed, using the selection of transgenic models for testing apoE-targeted therapeutics as an example (“Why”).