Brian J. Kirby

Despite advances in lipid-lowering treatment, atherosclerotic cardiovascular disease remains the leading cause of mortality, underscoring the need to address residual risk. Targeting both the synthesis and clearance of triglyceride (TG)-rich lipoproteins is a promising approach. Liver X receptor (LXR) repression can reduce plasma TG and cholesterol and improve insulin sensitivity by suppressing de novo lipogenesis and intestinal lipid absorption and enhancing clearance of TG-rich lipoproteins, but its clinical utility remains unexplored. Here we demonstrate the role of LXR inverse agonists in lipid metabolism and metabolic diseases in preclinical models and humans. Given concerns that systemic LXR repression may impair reverse cholesterol transport, we developed TLC-2716, an orally administered, gut- and liver-restricted LXR inverse agonist. In human liver organoids modeling steatohepatitis, TLC-2716 reduced lipid accumulation and suppressed inflammation and fibrotic gene expression. In a randomized, placebo-controlled phase 1 clinical trial, 14-day treatment with TLC-2716 was well tolerated (primary endpoints) and resulted in placebo-adjusted reductions up to 38.5% in plasma TG and 61% in postprandial remnant cholesterol (secondary endpoints). In conclusion, these results highlight the tolerability and therapeutic potential of TLC-2716 as a treatment for managing dyslipidemia and reducing residual atherosclerotic cardiovascular disease risk in humans. ClinicalTrials.gov identifier: NCT05483998 .

Background: Idiopathic calcium oxalate kidney stones are highly prevalent. There is need for evidence-based treatments that effectively reduce stone recurrence. LLX-424 is a prodrug of LLX-152, an inhibitor of glycolate oxidase (GO) in development for prevention of recurrent calcium oxalate stones. Oxalate is highly insoluble in the urine and can lead to formation of calcium oxalate stones. The majority of urinary oxalate is synthesized via GO in the liver. Inhibition of GO is expected to decrease urinary oxalate levels and recurrence of oxalate kidney stones. This Phase 1 study evaluated the safety, pharmacokinetics (PK) and pharmacodynamics (PD) of LLX-424 in healthy volunteers (HV). Methods: This randomized, double-blind, placebo-controlled study evaluated 5 single ascending dose (SAD) cohorts (50, 100, 200, 400, and 800 mg LLX-424) and 3 multiple ascending dose (MAD) cohorts (100, 200, and 300 mg LLX-424 once-daily for 14 days) during in-unit confinement. LLX-424 was administered fasted except for the 200 mg SAD cohort, in which a second dose was administered, after a washout, with a high-fat high-calorie meal, and the 300 mg MAD cohort which was administered under fed conditions. Plasma and urine concentrations of LLX-424, LLX-152, glycolate (the substrate of GO and a marker of target enzyme engagement) were determined by validated LC/MS/MS methods. Urine oxalate was assessed in MAD cohorts. Results: 72 subjects were randomized in the study. There were no serious or severe adverse events or adverse events requiring dose adjustment or discontinuation. LLX-424 and LLX-152 exhibited linear and time independent PK. A high-fat high-calorie meal modestly increased exposure of LLX-424 and LLX-152 (~55% Cmax and ~50-75% AUCinf). LLX-424 exhibited dose dependent increases in plasma glycolate PD parameters and reduced urinary oxalate. Conclusion: In this first-in-human study, LLX-424, an orally administered glycolate oxidase inhibitor, safely and effectively inhibited glycolate oxidase in a dose dependent manner; inhibition was maintained over an up to 2-week dosing period. The safety, tolerability, PK, and PD of LLX-424 supports further clinical evaluation in patients with recurrent kidney stones. Funding: Commercial Support - Lilac Therapeutics Inc.

Introduction and Objective: TLC-1180 is a novel, potent mitochondrial protonophore with extended pharmacology. Here, we evaluated ‘1180 as monotherapy (mono), in combination (combo) with semaglutide (SEMA), and for maintenance post SEMA discontinuation (D/C) in diet-induced obese (DIO) mice. Methods: Male DIO mice were treated with ‘1180 in 2 studies: de novo combo of ‘1180 + high-dose SEMA (SEMAhigh) followed by continued ‘1180 mono post D/C of SEMAhigh (4 + 2 wk maintenance), and ‘1180 mono (3 wk) followed by combo with SEMAlow (3 wk; sequencing). Body weight (BW) and fat (FM) and lean mass (LM) were quantified. Results: ‘1180 mono significantly reduced BW (-11-20%) and FM (-22-41%) without LM loss, while SEMAhigh mono reduced BW (-20%), FM (-36%), and LM (-8%). ‘1180 + SEMAhighde novo combo further lowered BW (-26%) and FM (-48%) vs SEMAhigh over 4 wks (Fig A). Post SEMAhigh D/C, ‘1180 maintenance blunted regain of BW and FM (+18%, +5%) vs vehicle (+46%, +27%) (Fig A). ’1180 + SEMAlow sequencing combo lowered BW and FM (–30%, -59%) vs SEMAhigh mono (-25%, -47%) (Fig B). In both studies, LM loss was similar between combo and SEMAhigh. Conclusion: In DIO mice, ‘1180 caused BW and FM loss while preserving LM. These benefits were amplified in combo with SEMA and maintained post SEMA D/C. These data highlight the potential of ‘1180 as mono and in combo with incretins in obesity and associated metabolic disorders. Disclosure M. Sharma: None. N. Sroda: Employee; OrsoBio, Inc. E. Murakami: Employee; OrsoBio, Inc. C. Logan: None. S. Weng: Employee; OrsoBio, Inc. B.J. Kirby: Employee; OrsoBio, Inc, Gossamer Bio, Inc. Consultant; Lilac Therapetutics, TERNS Pharmaceuticals, Actio Biosciences, SiteOne Therapeutics, IconOVir Bio, Recludix Pharma. R.P. Myers: Employee; OrsoBio, Inc. Stock/Shareholder; OrsoBio, Inc. M. Subramanian: Employee; OrsoBio, Inc. G.I. Shulman: Advisory Panel; Novo Nordisk. Consultant; Ionis Pharmaceuticals. Research Support; AstraZeneca, Merck & Co., Inc, ESPERION Therapeutics, Inc., Novo Nordisk. Advisory Panel; OrsoBio, Inc. A. Vijayakumar: Employee; OrsoBio, Inc.

Introduction and Objective: TLC-1180, a novel MP, and TLC-3595, an ACC2-selective inhibitor, enhance fatty acid oxidation via complementary mechanisms and are in development for obesity-associated disorders. Here, we compared the efficacy of a novel combination (combo) of ‘1180 and ‘3595 to semaglutide (SEMA) in diet-induced obese (DIO) mice. Methods: Male DIO mice housed at thermoneutrality were treated with ‘1180 or ‘3595 monotherapy (mono), a combo, or SEMA mono for 6 weeks. Body weight (BW) and fat and lean mass were quantified. Results: ‘3595 and ‘1180 mono lowered BW (-5% and -13%, respectively), primarily driven by fat mass loss (-12% and -27%, respectively) vs vehicle. ‘1180+’3595 combo further reduced BW by -16%, similar to SEMA (-17%, Fig A). Reductions in fat mass were similar with ‘1180+’3595 combo (-36%) and SEMA (-33%) (Fig B). While SEMA reduced food intake, no changes in food intake were seen with ‘1180+’3595 combo. Importantly, unlike SEMA which reduced lean mass, no decrease in lean mass was observed with ‘1180+’3595 combo (p<0.05 vs SEMA). Conclusion: In DIO mice, a novel combo of a MP and an ACC2 inhibitor caused comparable weight loss to an incretin but preserved lean mass. In sum, these data support the evaluation of combinations of these agents in people living with obesity and associated metabolic disorders. Disclosure N. Sroda: Employee; OrsoBio, Inc. M. Sharma: None. E. Murakami: Employee; OrsoBio, Inc. C. Logan: None. S. Weng: Employee; OrsoBio, Inc. B.J. Kirby: Employee; OrsoBio, Inc, Gossamer Bio, Inc. Consultant; Lilac Therapetutics, TERNS Pharmaceuticals, Actio Biosciences, SiteOne Therapeutics, IconOVir Bio, Recludix Pharma. R.P. Myers: Employee; OrsoBio, Inc. Stock/Shareholder; OrsoBio, Inc. M. Subramanian: Employee; OrsoBio, Inc. G.I. Shulman: Advisory Panel; Novo Nordisk. Consultant; Ionis Pharmaceuticals. Research Support; AstraZeneca, Merck & Co., Inc, ESPERION Therapeutics, Inc., Novo Nordisk. Advisory Panel; OrsoBio, Inc. A. Vijayakumar: Employee; OrsoBio, Inc.

Introduction and Objective: TLC-6740, a liver-targeted MP, regulates body weight via increased energy expenditure and is complementary to incretins that reduce food intake. We evaluated weight loss with a sequential combination of ‘6740 and semaglutide (SEMA), and ‘6740 maintenance therapy post SEMA discontinuation (D/C) in diet-induced obese (DIO) mice. Methods: Male DIO mice were treated with ‘6740 (6 wk), ‘6740 (3 wk) followed by combo with SEMAlow (3 wk), or ‘6740 post D/C of SEMAhigh (3 + 3 wk). Body weight (BW), fat (FM) and lean mass (LM), and oral glucose tolerance (oGTT) were assessed. Results: ‘6740 caused significant BW (-18%, 6 wk) and FM loss (-52%) without altering LM vs vehicle, while SEMAhigh reduced BW (-24%), FM (-67%), and LM (-16%). ‘6740+SEMAlow combo was well tolerated and further lowered BW, FM, and oGTT AUC (-33%, -77%, and -28%), but did not affect LM (Fig A). Post SEMAhigh D/C, ‘6740 maintained BW and FM loss similarly to mice continuing on SEMAhigh, unlike mice switched to vehicle (Fig B). ‘6740 maintenance also blunted the worsening of glucose tolerance post SEMAhigh D/C vs vehicle. Conclusion: In DIO mice, ‘6740 combo with SEMAlow was more efficacious than SEMAhigh, maintained weight loss post SEMA D/C, and was LM neutral. These data support evaluation of ‘6740 in combo with incretins in obesity and associated metabolic disorders. Disclosure N. Sroda: Employee; OrsoBio, Inc. M. Sharma: None. E. Murakami: Employee; OrsoBio, Inc. C. Logan: None. S. Weng: Employee; OrsoBio, Inc. B.J. Kirby: Employee; OrsoBio, Inc, Gossamer Bio, Inc. Consultant; Lilac Therapetutics, TERNS Pharmaceuticals, Actio Biosciences, SiteOne Therapeutics, IconOVir Bio, Recludix Pharma. R.P. Myers: Employee; OrsoBio, Inc. Stock/Shareholder; OrsoBio, Inc. M. Subramanian: Employee; OrsoBio, Inc. G.I. Shulman: Advisory Panel; Novo Nordisk. Consultant; Ionis Pharmaceuticals. Research Support; AstraZeneca, Merck & Co., Inc, ESPERION Therapeutics, Inc., Novo Nordisk. Advisory Panel; OrsoBio, Inc. A. Vijayakumar: Employee; OrsoBio, Inc.

Thrombosis, or pathological blood clot formation, is a dangerous and potentially fatal event that is often associated with implanted medical devices. Thrombi are comprised of platelets, plasma proteins, red blood cells (RBCs) and other molecular and cellular components. Extensive research has been conducted on the dynamics of platelet aggregation and deposition; however, the process of RBC entrapment remains under-explored. In this study, we used a microfluidic device and fluorescence microscopy to concurrently image platelets and RBCs during thrombus formation under flow and biomaterial conditions representative of implanted devices. We designed and applied MATLAB algorithms to track thrombi as they grew over time and interrogated thrombus growth and RBC accumulation trends. To probe the contributions of the unique mechanical properties of RBCs, we characterized the influence of induced RBC stiffness on thrombus forming behavior via imaging. To study the binding effects of known RBC surface antigens, we visualized thrombus formation with inhibition of glycoprotein (GP) IIb/IIIa and integrin-associated protein (IAP) respectively. We found that the accumulation and clustering of RBCs within the thrombi exhibited an inverse relationship with stiffness. Fibrillar structures were also present in these thrombi. Stiff RBCs enhanced platelet aggregate growth in all directions. Inhibition of surface binding proteins GPIIb/IIIa and IAP decreased the stability of thrombi. The results show that RBC deformability impacts entrapment location, fibril growth and spatial distribution. In turn, RBC binding stabilizes and anchors aggregates to surfaces. These findings will inform improved device design and multi-phase models of thrombosis that incorporate RBCs. STATEMENT OF SIGNIFICANCE: Dummy.

• In this randomized trial, colorectal cancer survivors achieved high adherence to a home-based moderate-intensity aerobic exercise prescription. • Exercise improved objective measures of fitness capacity and physical activity. • Exercise did not reduce inflammation in the overall sample; however, among the subgroup of participants with high inflammation at baseline, exercise had anti-inflammatory effects. • Among participants randomized to the exercise group, higher exercise adherence was correlated with a reduction in circulating tumor cells but not a reduction in circulating tumor DNA. The biological mechanisms by which postdiagnosis physical activity improves disease-free survival in colorectal cancer survivors remain incompletely understood. This trial tested the hypothesis that 12 weeks of moderate-intensity aerobic exercise, when compared with a control group, would change inflammation, circulating tumor cells (CTCs), and circulating tumor DNA (ctDNA) in a manner consistent with an improved cancer prognosis. This trial randomized Stages I–III colorectal cancer survivors to 12 weeks of home-based moderate-intensity aerobic exercise or a waitlist control group. The co-primary endpoints were high-sensitivity C-reactive protein (hs-CRP) and interleukin-6 (IL-6), secondary endpoints were soluble tumor necrosis factor-α receptor 2 (sTNFαR2) and CTCs, and the exploratory endpoint was tumor fraction quantified from ctDNA. Sixty subjects were randomized (age = 60.6 ± 10.8 years, mean ± SD; 39 (65%) females; 46 (77%) colonic primary tumor), and 59 (98%) subjects completed the study. Over 12 weeks, exercise adherence was 92% (95% confidence interval (95%CI): 86‒99). Exercise improved submaximal fitness capacity (0.36 metabolic equivalents; 95%CI: 0.05‒0.67; p = 0.025) and objectively measured moderate-to-vigorous-intensity physical activity (34.8%, 95%CI: 11.3‒63.1; p = 0.002) compared to control. Exercise did not change hs-CRP (20.9%, 95%CI: −17.1 to 76.2; p = 0.32), IL-6 (11.4%, 95%CI: −7.5 to 34.0; p = 0.25), or sTNFαR2 (−3.6%, 95%CI: −13.7 to 7.7; p = 0.52) compared to control. In the subgroup of subjects with elevated baseline hs-CRP ( n = 35, 58.3%), aerobic exercise reduced hs-CRP (−35.5%, 95%CI: −55.3 to −3.8; p = 0.031). Exercise did not change CTCs (0.59 cells/mL, 95%CI: −0.33 to 1.51; p = 0.21) or tumor fraction (0.0005, 95%CI: −0.0024 to 0.0034; p = 0.73). In exploratory analyses, higher aerobic exercise adherence correlated with a reduction in CTCs (ρ = −0.37, 95%CI: −0.66 to −0.08; p = 0.013). Colorectal cancer survivors achieved high adherence to a home-based moderate-intensity aerobic exercise prescription that improved fitness capacity and physical activity but did not reduce inflammation or change tumor endpoints from a liquid biopsy.

Metamaterials are composite structures whose extraordinary properties arise from a mesoscale organization of their constituents. Here, we introduce a different material class—viscosity metafluids. Specifically, we demonstrate that we can rapidly drive large viscosity oscillations in shear-thickened fluids using acoustic perturbations with kHz to MHz frequencies. Because the timescale for these oscillations can be orders of magnitude smaller than the timescales associated with the global material flow, we can construct metafluids whose resulting time-averaged viscosity is a composite of the thickened, high-viscosity and dethickened, low-viscosity states. We show that viscosity metafluids can be used to engineer a variety of unique properties including zero, infinite, and negative viscosities. The high degree of control over the resulting viscosity, the ease with which they can be accessed, and the variety of exotic properties achievable make viscosity metafluids attractive for uses in technologies ranging from coatings to cloaking to 3D printing. Published by the American Physical Society 2024

BACKGROUND Physical activity is associated with improved disease-free survival in colorectal cancer survivors. This report describes the purpose, design, recruitment, and exercise adherence results of the National Cancer Institute (NCI)-sponsored Exercise and Colorectal Cancer Treatment (EXACT) trial. METHODS The primary objective of the EXACT trial is to determine if randomization to 150 min per week of moderate-intensity aerobic exercise reduces systemic inflammation among stage I-III colorectal cancer survivors compared with a waitlist control group over 12 weeks. Participants were provided with an in-home treadmill and heart rate monitor. Characteristics associated with randomization using χ2 or Fisher's exact test for categorical variables and t-tests or analysis of covariance (ANCOVA). Exercise adherence was calculated as the total minutes exercised by total minutes prescribed. RESULTS Between August 2019 and February 2023, 3082 colorectal cancer survivors were invited to participate, 89 were screened, and 60 were randomized to the study protocol. Younger age (P = 0.02), female sex (P = 0.002), white race (P = 0.01), proximal time since tumor resection (P = 0.02), and regional tumor stage (P < 0.001) were associated with study participation. Average exercise adherence was 92.2 % (95 % CI: 85.5, 98.8) and all study participants achieved ≥80 % exercise adherence. Endpoint data collection was completed for all participants in May 2023. CONCLUSION The results from the EXACT trial will characterize the changes that occur from exercise to advance our understanding of the biological mechanisms by which exercise may prevent tumor recurrence and death in colorectal cancer survivors.

Background: Mitochondrial uncoupling is a validated approach to weight loss, however, toxicity due to systemic uncoupling has restricted clinical use. To enhance safety, TLC-6740 was developed as a liver-targeted protonophore (&amp;gt;40x liver/plasma ratio) for the treatment of obesity. Methods: In this randomized, double-blind Ph 1 study (NCT05822544), healthy subjects received single (SAD; n=48) or multiple ascending doses (MAD; n=60) of TLC-6740 (3-120 mg) PO once daily for 10 days (n=6-8 active vs n=2 placebo/group) while fasting. Eight subjects received TLC-6740 15 mg fasted and then fed to evaluate food effect. Results: In the MAD cohorts, the proportion of subjects with adverse events (AEs) was 58% with placebo and 50-88% with TLC-6740. All AEs were mild except 3 moderate AEs unrelated to treatment (anal fissure, 30 mg; heating pad burn, 120 mg; viral illness, placebo). No drug-related discontinuations, lab abnormalities, or temperature increases were observed. TLC-6740 had a median steady-state half-life of 18-33 hrs; AUC and Cmax increased in a dose-dependent manner with no impact of food or body weight. TLC-6740 caused significant, dose-dependent reductions in serum total and LDL-C (Figure). Conclusion: In this Ph 1 study, the liver-targeted protonophore TLC-6740 was well tolerated and led to dose-dependent improvements in lipids, supporting its potential for treatment of obesity. Disclosure E. Gane: None. R.S. Huss: None. J. Sur: Employee; OrsoBio, Inc. E. Murakami: Employee; OrsoBio, Inc. S. Weng: Employee; OrsoBio, Inc. B.J. Kirby: Consultant; OrsoBio, Inc. A. Shah: Consultant; OrsoBio, Inc. G.I. Shulman: None. M. Subramanian: Board Member; OrsoBio, Inc. A. Vijayakumar: Employee; OrsoBio, Inc. R.P. Myers: Employee; OrsoBio, Inc.

In this paper we report on the development of strategies used in an introductory fluid mechanics course that transitioned from a fully in-person mode of delivery to a hybrid setting.We describe two sets of instructional changes we used to support students' learning in the hybrid context: first, Matlab live script documents and second, "scavenger hunt" missions of finding, demonstrating, or building fluid mechanical systems in everyday life.We employ two different instruments to track students' experiences in this course.First, we compare students' performance in a fluid mechanics concept inventory assessment that they take at the end of each semester.In addition, we also adopt a set of items from the Motivated Strategies for Learning Questionnaire (MSLQ) to measure the impacts of these changes on students' motivations and attitudes.We reflect on the implications of this transition process and provide an outline of the future developments of this work.

Background: The liver X receptors are oxysterol-activated nuclear hormone receptors that regulate cholesterol homeostasis and de novo lipogenesis. TLC-2716 is an oral, liver-targeted inverse agonist of LXRα/β in development for severe hypertriglyceridemia (SHTG) and NASH. Methods: In this randomized, double-blind, placebo (PBO)-controlled Phase 1 study (NCT05483998), healthy subjects (n=8/2 TLC-2716 vs PBO/cohort) received single ascending doses (SAD) of TLC-2716 (0.5, 2, 6, 12, or 20 mg) or multiple ascending doses (MAD) QD for 14 days (0.5, 2, 6, or 12 mg) while fed or 6 mg fasted. Safety, PK, serum lipid parameters by NMR LipoProfile ® , and gene expression in PBMCs were evaluated. Results: The proportion of subjects with adverse events (AEs) was 60% with PBO and 13-50% with TLC-2716 in the SAD cohorts (n=50), and 40% and 13-88%, respectively, in the MAD cohorts (n=50). All AEs were mild except an unrelated, Grade 2 thrombophlebitis in a subject on TLC-2716 2 mg. TLC-2716 was rapidly absorbed (T max 2-4h) with a short steady-state half-life (T 1/2 ~1.5-2.5h) and low maximal plasma concentrations (C max &lt;7 ng/mL), consistent with rapid hepatic uptake. With fed dosing, TLC-2716 AUC and C max increased less than dose proportionally and were 25-50% lower on Day 14 vs Day 1. With fasted dosing, exposures were similar on Days 1 and 14 and ~3-fold higher than with fed dosing. Significant, dose-dependent reductions in serum triglycerides, total and LDL-C, total and small LDL particles, and TG/HDL-C ratio—but not HDL-C—were observed with TLC-2716 ( Figure ). Lipid reductions were greatest in subjects with higher baseline values. TLC-2716 did not impact LXR target gene expression (e.g., reverse cholesterol transporters ABCA1 , ABCG1 ) in PBMCs, supporting its liver-targeted profile. Conclusion: The liver-targeted LXR inverse agonist TLC-2716 was well tolerated and caused significant lipid lowering in healthy subjects, supporting its evaluation in patients with SHTG and NASH.

Background: HER2-low is a newly defined category with HER2 1+ or 2+ expression by immunohistochemistry (IHC) and lack of HER2 gene amplification measured by in situ hybridization (ISH). Much remains unknown about the HER2-low status across tumor types and changes in HER2 status between primary and metastatic samples. Methods: HER2 expression by IHC was evaluated in 4,701 patients with solid tumors. We have evaluated the HER2 expression by IHC and amplification by FISH in paired breast and gastric/gastroesophageal (GEJ) primary and metastatic samples. HER2 expression was correlated with ERBB2 genomic alterations evaluated by next-generation sequencing (NGS) in non-breast, non-gastric/GEJ samples. Results: HER2 expression (HER2 IHC 1–3+) was found in half (49.8%) of cancers, with HER2-low (1 or 2+) found in many tumor types: 47.1% in breast, 34.6% in gastric/GEJ, 50.0% in salivary gland, 46.9% in lung, 46.5% in endometrial, 46% in urothelial and 45.5 % of gallbladder cancers. The concordance evaluation of HER2 expression between primary and metastatic breast cancer samples, showed that HER2 3+ remained unchanged in 87.1% with a strong agreement between primary and metastatic samples, with a weighted kappa (K) of 0.85 (95% CI, 0.79–0.91). ERBB2 alterations were identified in 117 (7.5%) patients with non-breast, non-gastric/GEJ solid tumors who had NGS testing. Of 1,436 patients without ERBB2 alterations, 512 (35.7%) showed any level HER2 expression by IHC. Conclusion: Our results show that HER2-low expression is frequently found across tumor types. These findings suggest that many patients with HER2-low solid tumors might benefit from HER2-targeted therapies.

&lt;div&gt;AbstractPurpose:&lt;p&gt;Biomarkers aiding treatment optimization in metastatic castration-resistant prostate cancer (mCRPC) are scarce. The presence or absence of androgen receptor (AR) splice variants, AR-V7 and AR&lt;sup&gt;v567es&lt;/sup&gt;, in mCRPC patient circulating tumor cells (CTC) may be associated with taxane treatment outcomes.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Experimental Design:&lt;/b&gt; A novel digital droplet PCR (ddPCR) assay assessed AR-splice variant expression in CTCs from patients receiving docetaxel or cabazitaxel in TAXYNERGY (NCT01718353). Patient outcomes were examined according to AR-splice variant expression, including prostate-specific antigen (PSA)&lt;sub&gt;50&lt;/sub&gt; response and progression-free survival (PFS).&lt;/p&gt;Results:&lt;p&gt;Of the 54 evaluable patients, 36 (67%) were AR-V7&lt;sup&gt;+&lt;/sup&gt;, 42 (78%) were AR&lt;sup&gt;v567es+&lt;/sup&gt;, 29 (54%) were double positive, and 5 (9%) were double negative. PSA&lt;sub&gt;50&lt;/sub&gt; response rates at any time were numerically higher for AR-V7&lt;sup&gt;−&lt;/sup&gt; versus AR-V7&lt;sup&gt;+&lt;/sup&gt; (78% vs. 58%; &lt;i&gt;P&lt;/i&gt; = 0.23) and for AR&lt;sup&gt;v567es−&lt;/sup&gt; versus AR&lt;sup&gt;v567es+&lt;/sup&gt; (92% vs. 57%; &lt;i&gt;P&lt;/i&gt; = 0.04) patients. When AR-V mRNA status was correlated with change in nuclear AR from cycle 1 day 1 to day 8 (&lt;i&gt;n&lt;/i&gt; = 24), AR-V7&lt;sup&gt;+&lt;/sup&gt; patients (&lt;i&gt;n&lt;/i&gt; = 16) had a 0.4% decrease versus a 12.9% and 26.7% decrease in AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es−&lt;/sup&gt; (&lt;i&gt;n&lt;/i&gt; = 3) and AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es+&lt;/sup&gt; (&lt;i&gt;n&lt;/i&gt; = 5) patients, respectively, suggesting a dominant role for AR-V7 over AR&lt;sup&gt;v567es&lt;/sup&gt;. Median PFS was 12.02 versus 8.48 months for AR-V7&lt;sup&gt;−&lt;/sup&gt; versus AR-V7&lt;sup&gt;+&lt;/sup&gt; (HR = 0.38; &lt;i&gt;P&lt;/i&gt; = 0.01), and 12.71 versus 7.29 months for AR&lt;sup&gt;v567es−&lt;/sup&gt; versus AR&lt;sup&gt;v567es+&lt;/sup&gt; (HR = 0.37; &lt;i&gt;P&lt;/i&gt; = 0.02). For AR-V7&lt;sup&gt;+&lt;/sup&gt;, AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es+&lt;/sup&gt;, and AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es−&lt;/sup&gt; patients, median PFS was 8.48, 11.17, and 16.62 months, respectively (&lt;i&gt;P&lt;/i&gt; = 0.0013 for trend).&lt;/p&gt;Conclusions:&lt;p&gt;Although detection of both CTC-specific AR-V7 and AR&lt;sup&gt;v567es&lt;/sup&gt; by ddPCR influenced taxane outcomes, AR-V7 primarily mediated the prognostic impact. The absence of both variants was associated with the best response and PFS with taxane treatment.&lt;/p&gt;&lt;p&gt;See related commentary by Dehm et al., p. 1696&lt;/p&gt;&lt;/div&gt;

&lt;p&gt;Visual representation of AR-FL and AR splice variant expression per patient at baseline Data are shown as positive (green for AR-FL; red for AR-V7; blue for ARv567es) or negative (white boxes) for each transcript. AR, androgen receptor; FL, full length&lt;/p&gt;

&lt;p&gt;Assay sensitivity in 22RV1 prostate cancer cells spiked-in healthy donor blood run through the GEDI device. AR-FL (blue) and AR-V7 (red) mRNA expression (#copies per sample) was determined by ddPCR in varying amounts of 22RV1 cells in the presence of healthy donor blood run through the GEDI device. The assay, reliably and reproducibly detects both transcripts in single spiked-in cells. The table below the graph shows the raw data (copy number) for each transcript per condition. Healthy donor blood PBMCs alone were used as a control. AR, androgen receptor; ddPCR, droplet digital polymerase chain reaction; FL, full length; GEDI, geometrically enhanced differential immunocapture; PBMC, peripheral blood mononuclear cell.&lt;/p&gt;

&lt;p&gt;ddPCR primer and probe sequences AR, androgen receptor; FL, full length.&lt;/p&gt;

&lt;p&gt;Copy number for AR-FL, AR-V7 and ARv567es for the evaluable population at baseline AR, androgen receptor; FL, full length.&lt;/p&gt;

&lt;p&gt;AR-FL and AR-V expression in healthy donor blood run through the GEDI device. The mRNA expression levels of AR-FL (blue), AR-v7 (red) and AR-v567es (green) were analyzed in healthy donor blood from 10 volunteers. As previously described, 1 mL of healthy donor peripheral blood was processed through the GEDI microfluidic device. The table below the graph shows the raw data (copy number) for each transcript per sample. AR, androgen receptor; ddPCR, droplet digital polymerase chain reaction; FL, full length; GEDI, geometrically enhanced differential immunocapture; mRNA, messenger ribonucleic acid.&lt;/p&gt;

&lt;p&gt;Confirmation of ARv567es amplicon by Sanger Sequencing A) Agarose gel electrophoresis showing the PCR-amplified ARv567es amplicon using ddPCR primers. Lane 1 DNA ladder from New England Biolabs, Inc; Lane 2 HEK293 cells transfected with Empty Vector (Negative control); Lane 3 HEK293 cells transfected with ARv567es-variant plasmid (Positive control); Lane 4 HEK293 cells transfected with ARv567es-variant plasmid, but with no reverse transcriptase (Negative control); Lanes 5-6 is CTC-derived DNA from patients #7 and #4 respectively. The red arrow depicts the expected size of ARv567es amplicon. B) Representative DNA chromatogram following Sanger Sequencing with the reverse and forward primers for the ARv567es amplicon (Size = 95 bps). Data shown for patient #7 amplicon.&lt;/p&gt;

&lt;p&gt;Copy number for AR-V7 and ARv567es stratified by PSA response AR, androgen receptor; IQ, interquartile; PSA50, 50% reduction from baseline in prostate-specific antigen; SD, standard deviation.&lt;/p&gt;

&lt;div&gt;AbstractPurpose:&lt;p&gt;Biomarkers aiding treatment optimization in metastatic castration-resistant prostate cancer (mCRPC) are scarce. The presence or absence of androgen receptor (AR) splice variants, AR-V7 and AR&lt;sup&gt;v567es&lt;/sup&gt;, in mCRPC patient circulating tumor cells (CTC) may be associated with taxane treatment outcomes.&lt;/p&gt;&lt;p&gt;&lt;b&gt;Experimental Design:&lt;/b&gt; A novel digital droplet PCR (ddPCR) assay assessed AR-splice variant expression in CTCs from patients receiving docetaxel or cabazitaxel in TAXYNERGY (NCT01718353). Patient outcomes were examined according to AR-splice variant expression, including prostate-specific antigen (PSA)&lt;sub&gt;50&lt;/sub&gt; response and progression-free survival (PFS).&lt;/p&gt;Results:&lt;p&gt;Of the 54 evaluable patients, 36 (67%) were AR-V7&lt;sup&gt;+&lt;/sup&gt;, 42 (78%) were AR&lt;sup&gt;v567es+&lt;/sup&gt;, 29 (54%) were double positive, and 5 (9%) were double negative. PSA&lt;sub&gt;50&lt;/sub&gt; response rates at any time were numerically higher for AR-V7&lt;sup&gt;−&lt;/sup&gt; versus AR-V7&lt;sup&gt;+&lt;/sup&gt; (78% vs. 58%; &lt;i&gt;P&lt;/i&gt; = 0.23) and for AR&lt;sup&gt;v567es−&lt;/sup&gt; versus AR&lt;sup&gt;v567es+&lt;/sup&gt; (92% vs. 57%; &lt;i&gt;P&lt;/i&gt; = 0.04) patients. When AR-V mRNA status was correlated with change in nuclear AR from cycle 1 day 1 to day 8 (&lt;i&gt;n&lt;/i&gt; = 24), AR-V7&lt;sup&gt;+&lt;/sup&gt; patients (&lt;i&gt;n&lt;/i&gt; = 16) had a 0.4% decrease versus a 12.9% and 26.7% decrease in AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es−&lt;/sup&gt; (&lt;i&gt;n&lt;/i&gt; = 3) and AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es+&lt;/sup&gt; (&lt;i&gt;n&lt;/i&gt; = 5) patients, respectively, suggesting a dominant role for AR-V7 over AR&lt;sup&gt;v567es&lt;/sup&gt;. Median PFS was 12.02 versus 8.48 months for AR-V7&lt;sup&gt;−&lt;/sup&gt; versus AR-V7&lt;sup&gt;+&lt;/sup&gt; (HR = 0.38; &lt;i&gt;P&lt;/i&gt; = 0.01), and 12.71 versus 7.29 months for AR&lt;sup&gt;v567es−&lt;/sup&gt; versus AR&lt;sup&gt;v567es+&lt;/sup&gt; (HR = 0.37; &lt;i&gt;P&lt;/i&gt; = 0.02). For AR-V7&lt;sup&gt;+&lt;/sup&gt;, AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es+&lt;/sup&gt;, and AR-V7&lt;sup&gt;−&lt;/sup&gt;/AR&lt;sup&gt;v567es−&lt;/sup&gt; patients, median PFS was 8.48, 11.17, and 16.62 months, respectively (&lt;i&gt;P&lt;/i&gt; = 0.0013 for trend).&lt;/p&gt;Conclusions:&lt;p&gt;Although detection of both CTC-specific AR-V7 and AR&lt;sup&gt;v567es&lt;/sup&gt; by ddPCR influenced taxane outcomes, AR-V7 primarily mediated the prognostic impact. The absence of both variants was associated with the best response and PFS with taxane treatment.&lt;/p&gt;&lt;p&gt;See related commentary by Dehm et al., p. 1696&lt;/p&gt;&lt;/div&gt;

&lt;p&gt;AR-FL and AR-V expression in healthy donor blood run through the GEDI device. The mRNA expression levels of AR-FL (blue), AR-v7 (red) and AR-v567es (green) were analyzed in healthy donor blood from 10 volunteers. As previously described, 1 mL of healthy donor peripheral blood was processed through the GEDI microfluidic device. The table below the graph shows the raw data (copy number) for each transcript per sample. AR, androgen receptor; ddPCR, droplet digital polymerase chain reaction; FL, full length; GEDI, geometrically enhanced differential immunocapture; mRNA, messenger ribonucleic acid.&lt;/p&gt;

&lt;p&gt;Assay sensitivity in 22RV1 prostate cancer cells spiked-in healthy donor blood run through the GEDI device. AR-FL (blue) and AR-V7 (red) mRNA expression (#copies per sample) was determined by ddPCR in varying amounts of 22RV1 cells in the presence of healthy donor blood run through the GEDI device. The assay, reliably and reproducibly detects both transcripts in single spiked-in cells. The table below the graph shows the raw data (copy number) for each transcript per condition. Healthy donor blood PBMCs alone were used as a control. AR, androgen receptor; ddPCR, droplet digital polymerase chain reaction; FL, full length; GEDI, geometrically enhanced differential immunocapture; PBMC, peripheral blood mononuclear cell.&lt;/p&gt;

&lt;p&gt;Visual representation of AR-FL and AR splice variant expression per patient at baseline Data are shown as positive (green for AR-FL; red for AR-V7; blue for ARv567es) or negative (white boxes) for each transcript. AR, androgen receptor; FL, full length&lt;/p&gt;

&lt;p&gt;Copy number for AR-V7 and ARv567es stratified by PSA response AR, androgen receptor; IQ, interquartile; PSA50, 50% reduction from baseline in prostate-specific antigen; SD, standard deviation.&lt;/p&gt;

&lt;p&gt;ddPCR primer and probe sequences AR, androgen receptor; FL, full length.&lt;/p&gt;

&lt;p&gt;Copy number for AR-FL, AR-V7 and ARv567es for the evaluable population at baseline AR, androgen receptor; FL, full length.&lt;/p&gt;

&lt;p&gt;Confirmation of ARv567es amplicon by Sanger Sequencing A) Agarose gel electrophoresis showing the PCR-amplified ARv567es amplicon using ddPCR primers. Lane 1 DNA ladder from New England Biolabs, Inc; Lane 2 HEK293 cells transfected with Empty Vector (Negative control); Lane 3 HEK293 cells transfected with ARv567es-variant plasmid (Positive control); Lane 4 HEK293 cells transfected with ARv567es-variant plasmid, but with no reverse transcriptase (Negative control); Lanes 5-6 is CTC-derived DNA from patients #7 and #4 respectively. The red arrow depicts the expected size of ARv567es amplicon. B) Representative DNA chromatogram following Sanger Sequencing with the reverse and forward primers for the ARv567es amplicon (Size = 95 bps). Data shown for patient #7 amplicon.&lt;/p&gt;

BACKGROUND AND OBJECTIVE: Cilofexor is a selective farnesoid X receptor (FXR) agonist in development for the treatment of nonalcoholic steatohepatitis and primary sclerosing cholangitis. Our objective was to evaluate potential drug-drug interactions of cilofexor as a victim and as a perpetrator. METHODS: In this Phase 1 study, healthy adult participants (n = 18-24 per each of the 6 cohorts) were administered cilofexor in combination with either perpetrators or substrates of cytochrome P-450 (CYP) enzymes and drug transporters. RESULTS: In total, 131 participants completed the study. As a victim, cilofexor area under the curve (AUC) was 651%, 795%, and 175% when administered following single-dose cyclosporine (600 mg; organic anion transporting polypeptide [OATP]/P-glycoprotein [P-gp]/CYP3A inhibitor), single-dose rifampin (600 mg; OATP1B1/1B3 inhibitor), and multiple-dose gemfibrozil (600 mg twice daily [BID]; CYP2C8 inhibitor), respectively, compared with the administration of cilofexor alone. Cilofexor AUC was 33% when administered following multiple-dose rifampin (600 mg; OATP/CYP/P-gp inducer). Multiple-dose voriconazole (200 mg BID; CYP3A4 inhibitor) and grapefruit juice (16 ounces; intestinal OATP inhibitor) did not affect cilofexor exposure. As a perpetrator, multiple-dose cilofexor did not affect the exposure of midazolam (2 mg; CYP3A substrate), pravastatin (40 mg; OATP substrate), or dabigatran etexilate (75 mg; intestinal P-gp substrate), but atorvastatin (10 mg; OATP/CYP3A4 substrate) AUC was 139% compared with atorvastatin administered alone. CONCLUSION: Cilofexor may be coadministered with inhibitors of P-gp, CYP3A4, or CYP2C8 without the need for dose modification. Cilofexor may be coadministered with OATP, BCRP, P-gp, and/or CYP3A4 substrates-including statins-without dose modification. However, coadministration of cilofexor with strong hepatic OATP inhibitors, or with strong or moderate inducers of OATP/CYP2C8, is not recommended.

A microvalve having a cast-in-place and lithographically shaped mobile, polymer monolith for fluid flow control in microfluidic devices and method of manufacture. The microvalve contains a porous fluorinated polymer monolithic element whose pores are filled with an electrically insulating, high dielectric strength fluid, typically a perfluorinated liquid. This combination provides a microvalve that combines high dielectric strength with extremely low electrical conductivity. These microvalves have been shown to have resistivities of at least 100 G.OMEGA. and are compatible with solvents such as water at a pH between 2.7 and 9.0, 1-1 propanol, acetonitrile, and acetone.