Classification of Features across Five CURE Networks Reveals Opportunities to Improve Course Design, Instruction, and Equity.

Course-based undergraduate research experiences (CUREs) are tools used to introduce students to authentic participation in science. Several specific CUREs have been shown to benefit students' interest and retention in the biological sciences. Nevertheless, CUREs vary greatly in terms of their context, methodology, and degree of research authenticity, so different types of CUREs may differently influence student outcomes. This programmatic diversity poses a challenge to educators who want to better understand which course components and features are reliably present in a CURE curriculum. To address these issues, we identified, catalogued, and classified 112 potential features of CUREs across the biosciences. To develop the list, we interviewed instructors experienced with teaching individual and large networked CUREs across a diversity of the biological disciplines, including: Squirrel-Net (field-based animal behavior), SEA-PHAGES (wet lab microbiology and computational microbiology), Tiny Earth (environmental and wet lab microbiology), PARE (environmental microbiology), and the Genomics Education Partnership (eukaryotic computational biology). Twenty-five interviewees contributed expert content in terms of CURE features and classification of those items into an organized list. The resulting list's categories encompasses student experiences with the following: (i) the scientific process; (ii) technical aspects of science; (iii) the professional development associated with research; and (iv) building scientific identity. The most striking insight was that CUREs vary widely in terms of which features they contain, since different CUREs will by necessity have different approaches to science and student involvement. We also identified several features commonly thought to be crucial to CUREs yet have ambiguous definitions. This ambiguity can potentially confound efforts to make CUREs research-authentic and aligned with the central goals of science. We disambiguate these terms and represent their varied meanings throughout the classification. We also provide instructor-friendly supplementary worksheets along with considerations for instructors interested in expanding their CURE course design, instruction, and equity.

Broadening Access to STEM through the Community College: Investigating the Role of Course-Based Research Experiences (CREs).

Broadening access to science, technology, engineering, and mathematics (STEM) professions through the provision of early-career research experiences for a wide range of demographic groups is important for the diversification of the STEM workforce. The size and diversity of the community college system make it a prime educational site for achieving this aim. However, some evidence shows that women and Black, Latinx, and Native American student groups have been hindered in STEM at the community college level. One option for enhancing persistence in STEM is to incorporate the course-based research experiences (CREs) into the curriculum as a replacement for the prevalent traditional laboratory. This can be achieved through the integration of community colleges within extant, multi-institutional CREs such as the SEA-PHAGES program. Using a propensity score-matching technique, students in a CRE and traditional laboratory were compared on a range of psychosocial variables (project ownership, self-efficacy, science identity, scientific community values, and networking). Results revealed higher ratings for women and persons excluded because of their ethnicity or race (PEERs) in the SEA-PHAGES program on important predictors of persistence such as project ownership and science identity. This suggests that the usage of CREs at community colleges could have positive effects in addressing the gender gap for women and enhance inclusiveness for PEER students in STEM.

Insulin Prevents Hypercholesterolemia by Suppressing 12α-Hydroxylated Bile Acids.

BACKGROUND: The risk of cardiovascular disease in type 1 diabetes remains extremely high, despite marked advances in blood glucose control and even the widespread use of cholesterol synthesis inhibitors. Thus, a deeper understanding of insulin regulation of cholesterol metabolism, and its disruption in type 1 diabetes, could reveal better treatment strategies. METHODS: To define the mechanisms by which insulin controls plasma cholesterol levels, we knocked down the insulin receptor, FoxO1, and the key bile acid synthesis enzyme, CYP8B1. We measured bile acid composition, cholesterol absorption, and plasma cholesterol. In parallel, we measured markers of cholesterol absorption and synthesis in humans with type 1 diabetes treated with ezetimibe and simvastatin in a double-blind crossover study. RESULTS: Mice with hepatic deletion of the insulin receptor showed marked increases in 12α-hydroxylated bile acids, cholesterol absorption, and plasma cholesterol. This phenotype was entirely reversed by hepatic deletion of FoxO1. FoxO1 is inhibited by insulin and required for the production of 12α-hydroxylated bile acids, which promote intestinal cholesterol absorption and suppress hepatic cholesterol synthesis. Knockdown of Cyp8b1 normalized 12α-hydroxylated bile acid levels and completely prevented hypercholesterolemia in mice with hepatic deletion of the insulin receptor (n=5-30), as well as mouse models of type 1 diabetes (n=5-22). In parallel, the cholesterol absorption inhibitor, ezetimibe, normalized cholesterol absorption and low-density lipoprotein cholesterol in patients with type 1 diabetes as well as, or better than, the cholesterol synthesis inhibitor, simvastatin (n=20). CONCLUSIONS: Insulin, by inhibiting FoxO1 in the liver, reduces 12α-hydroxylated bile acids, cholesterol absorption, and plasma cholesterol levels. Thus, type 1 diabetes leads to a unique set of derangements in cholesterol metabolism, with increased absorption rather than synthesis. These derangements are reversed by ezetimibe, but not statins, which are currently the first line of lipid-lowering treatment in type 1 diabetes. Taken together, these data suggest that a personalized approach to lipid lowering in type 1 diabetes may be more effective and highlight the need for further studies specifically in this group of patients.

Instructional Models for Course-Based Research Experience (CRE) Teaching.

The course-based research experience (CRE) with its documented educational benefits is increasingly being implemented in science, technology, engineering, and mathematics education. This article reports on a study that was done over a period of 3 years to explicate the instructional processes involved in teaching an undergraduate CRE. One hundred and two instructors from the established and large multi-institutional SEA-PHAGES program were surveyed for their understanding of the aims and practices of CRE teaching. This was followed by large-scale feedback sessions with the cohort of instructors at the annual SEA Faculty Meeting and subsequently with a small focus group of expert CRE instructors. Using a qualitative content analysis approach, the survey data were analyzed for the aims of inquiry instruction and pedagogical practices used to achieve these goals. The results characterize CRE inquiry teaching as involving three instructional models: 1) being a scientist and generating data; 2) teaching procedural knowledge; and 3) fostering project ownership. Each of these models is explicated and visualized in terms of the specific pedagogical practices and their relationships. The models present a complex picture of the ways in which CRE instruction is conducted on a daily basis and can inform instructors and institutions new to CRE teaching.

Bridging Trade-Offs between Traditional and Course-Based Undergraduate Research Experiences by Building Student Communication Skills, Identity, and Interest.

Undergraduate research plays an important role in the development of science students. The two most common forms of undergraduate research are those in traditional settings (such as internships and research-for-credit in academic research labs) and course-based undergraduate research experiences (CUREs). Both of these settings offer many benefits to students, yet they have unique strengths and weaknesses that lead to trade-offs. Traditional undergraduate research experiences (UREs) offer the benefits of personalized mentorship and experience in a professional setting, which help build students' professional communication skills, interest, and scientific identity. However, UREs can reach only a limited number of students. On the other end of the trade-off, CUREs offer research authenticity in a many-to-one classroom research environment that reaches more students. CUREs provide real research experience in a collaborative context, but CUREs are not yet necessarily equipping students with all of the experiences needed to transition into a research lab environment outside the classroom. We propose that CURE instructors can bridge trade-offs between UREs and CUREs by deliberately including learning goals and activities in CUREs that recreate the benefits of UREs, specifically in the areas of professional communication, scientific identify, and student interest. To help instructors implement this approach, we provide experience- and evidence-based guidance for student-centered, collaborative learning opportunities.

A Framework of College Student Buy-in to Evidence-Based Teaching Practices in STEM: The Roles of Trust and Growth Mindset.

Evidence-based teaching practices (EBPs) foster college science, technology, engineering, and mathematics (STEM) students' engagement and performance, yet our knowledge of what contributes to the effectiveness of these practices is less established. We propose a framework that links four social-cognitive variables-students' trust in their instructors, growth mindset, buy-in to instructional practices, and course engagement-to long-standing desired student outcomes of academic performance and intent to persist in science. This framework was tested in classrooms identified as having a high level of EBP implementation with a multi-institutional sample of 2102 undergraduates taught by 14 faculty members. Results indicate that the buy-in framework is a valid representation of college students' learning experiences within EBP contexts overall as well as across underrepresented student groups. In comparison to students' level of growth mindset, students' trust in their instructors was more than twice as predictive of buy-in to how the course was being taught, suggesting that students' views of their instructors are more associated with thriving in a high-EBP course environment than their views of intelligence. This study contributes to the dialogue on transforming undergraduate STEM education by providing a validated student buy-in framework as a lens to understand how EBPs enhance student outcomes.

Using Pathway Modeling to Evaluate and Improve Student-Centered Teaching Practices in Co-Taught College Science Courses.

Student-centered teaching practices such as active learning continue to gain momentum in college science education. Many instructors committed to these innovative practices transform their classrooms beyond the standard lecture. Nevertheless, widespread implementation of these practices is limited, because the learning benefits for students are often attained through increased instructional complexity to which many instructors cannot commit. When co-instructors are teaching the course, the level of commitment to building a student-centered classroom may be even more profound. For these reasons, new tools are needed to help instructors and co-instructors plan, organize, evaluate, and communicate their classroom innovations. Pathway modeling is a tool with the potential to fill this gap. Unlike curriculum mapping-which identifies academic content gaps, redundancies, and misalignments by examining a series of courses within a plan of study-course pathway modeling creates a visual map of a single course and reveals how teaching practices influence short-, mid-, and long-term student learning outcomes. This essay demonstrates how course pathway modeling can help co-instructors better represent the complexity of student-centered teaching practices. We include guides for creating course pathway models and discuss how this approach offers the potential to improve curricular design, course evaluation, student assessment, and communication between co-instructors.

Reversal of hypertriglyceridemia in diabetic BTBR ob/ob mice does not prevent nephropathy.

The etiology of diabetic nephropathy in type 2 diabetes is multifactorial. Sustained hyperglycemia is a major contributor, but additional contributions come from the hypertension, obesity, and hyperlipidemia that are also commonly present in patients with type 2 diabetes and nephropathy. The leptin deficient BTBR ob/ob mouse is a model of type 2 diabetic nephropathy in which hyperglycemia, obesity, and hyperlipidemia, but not hypertension, are present. We have shown that reversal of the constellation of these metabolic abnormalities with leptin replacement can reverse the morphologic and functional manifestations of diabetic nephropathy. Here we tested the hypothesis that reversal specifically of the hypertriglyceridemia, using an antisense oligonucleotide directed against ApoC-III, an apolipoprotein that regulates the interactions of VLDL (very low density lipoproteins) with the LDL receptor, is sufficient to ameliorate the nephropathy of Type 2 diabetes. Antisense treatment resulted in reduction of circulating ApoC-III protein levels and resulted in substantial lowering of triglycerides to near-normal levels in diabetic mice versus controls. Antisense treatment did not ameliorate proteinuria or pathologic manifestations of diabetic nephropathy, including podocyte loss. These studies indicate that pathologic manifestations of diabetic nephropathy are unlikely to be reduced by lipid-lowering therapeutics alone, but does not preclude a role for such interventions to be used in conjunction with other therapeutics commonly employed in the treatment of diabetes and its complications.

Monoacylglycerol Acyltransferase 1 Knockdown Exacerbates Hepatic Ischemia/Reperfusion Injury in Mice With Hepatic Steatosis.

Nonalcoholic fatty liver disease (NAFLD) is becoming the most common indication for liver transplantation. The growing prevalence of NAFLD not only increases the demand for liver transplantation, but it also limits the supply of available organs because steatosis predisposes grafts to ischemia/reperfusion injury (IRI) and many steatotic grafts are discarded. We have shown that monoacylglycerol acyltransferase (MGAT) 1, an enzyme that converts monoacylglycerol to diacylglycerol, is highly induced in animal models and patients with NAFLD and is an important mediator in NAFLD-related insulin resistance. Herein, we sought to determine whether Mogat1 (the gene encoding MGAT1) knockdown in mice with hepatic steatosis would reduce liver injury and improve liver regeneration following experimental IRI. Antisense oligonucleotides (ASO) were used to knockdown the expression of Mogat1 in a mouse model of NAFLD. Mice then underwent surgery to induce IRI. We found that Mogat1 knockdown reduced hepatic triacylglycerol accumulation, but it unexpectedly exacerbated liver injury and mortality following experimental ischemia/reperfusion surgery in mice on a high-fat diet. The increased liver injury was associated with robust effects on the hepatic transcriptome following IRI including enhanced expression of proinflammatory cytokines and chemokines and suppression of enzymes involved in intermediary metabolism. These transcriptional changes were accompanied by increased signs of oxidative stress and an impaired regenerative response. We have shown that Mogat1 knockdown in a mouse model of NAFLD exacerbates IRI and inflammation and prolongs injury resolution, suggesting that Mogat1 may be necessary for liver regeneration following IRI and that targeting this metabolic enzyme will not be an effective treatment to reduce steatosis-associated graft dysfunction or failure.

Benefits of a College STEM Faculty Development Initiative: Instructors Report Increased and Sustained Implementation of Research-Based Instructional Strategies.

The Summer Institutes on Scientific Teaching (SI) is a faculty development workshop in which science, technology, engineering, and mathematics (STEM) instructors, particularly from biology, are trained in the Scientific Teaching (ST) pedagogy. While participants have generally reported positive experiences, we aimed to assess how the SI affected participants' teaching practices. Building on a previously developed taxonomy of ST practices, we surveyed SI participants from the 2004-2014 SI classes regarding specific ST practices. Participants' self-reported use and implementation of ST practices increased immediately after SI attendance as well as over a longer time frame, suggesting that implementation persisted and even increased with time. However, instructors reported implementation gains for some practices more than others. The practices with the highest gains were engaging students in their own learning, using learning goals in course design, employing formative assessment, developing overarching course learning goals, representing science as a process, and facilitating group discussion activities. We propose that the ST practices showing the greatest gains may serve as beneficial focal points for professional development programs, while practices with smaller gains may require modified dissemination approaches or support structures.

Communicating Complex STEM Program Evaluation to Diverse Stakeholders.

Visual representations, such as pathway models, are increasingly being used to both communicate higher education science, technology, engineering, and mathematics (STEM) education program evaluation plans as well as accurately represent complex programs and the systems within which the educational programs reside. However, these representations can be overwhelming to audiences that are not familiar with the program's structure or engaged in the evaluation process. The goal of this methods essay is to help both evaluators and discipline-based education researchers improve communication about program evaluation with a variety of stakeholders. We propose a three-stage method for developing progressively less complex visualizations to build affordances that help make the program evaluation process and statements of program impact more meaningful to a wider range of audiences. The creation of less complex visualizations can facilitate understanding by allowing a stakeholder to more easily "see" the structure of the program and thereby may evoke a greater willingness to take action and make meaningful programmatic changes based on strategic evaluation planning. To aid readers, we describe how we modified the Systems Evaluation Protocol (SEP) to develop simplified visualizations when evaluating a long-standing college science faculty development program, the Summer Institutes on Scientific Teaching.

Lipid Droplet-Derived Monounsaturated Fatty Acids Traffic via PLIN5 to Allosterically Activate SIRT1.

Lipid droplets (LDs) provide a reservoir for triacylglycerol storage and are a central hub for fatty acid trafficking and signaling in cells. Lipolysis promotes mitochondrial biogenesis and oxidative metabolism via a SIRT1/PGC-1α/PPARα-dependent pathway through an unknown mechanism. Herein, we identify that monounsaturated fatty acids (MUFAs) allosterically activate SIRT1 toward select peptide-substrates such as PGC-1α. MUFAs enhance PGC-1α/PPARα signaling and promote oxidative metabolism in cells and animal models in a SIRT1-dependent manner. Moreover, we characterize the LD protein perilipin 5 (PLIN5), which is known to enhance mitochondrial biogenesis and function, to be a fatty-acid-binding protein that preferentially binds LD-derived monounsaturated fatty acids and traffics them to the nucleus following cAMP/PKA-mediated lipolytic stimulation. Thus, these studies identify the first-known endogenous allosteric modulators of SIRT1 and characterize a LD-nuclear signaling axis that underlies the known metabolic benefits of MUFAs and PLIN5.

Obesity-linked suppression of membrane-bound O-acyltransferase 7 (MBOAT7) drives non-alcoholic fatty liver disease.

Recent studies have identified a genetic variant rs641738 near two genes encoding membrane bound O-acyltransferase domain-containing 7 (MBOAT7) and transmembrane channel-like 4 (TMC4) that associate with increased risk of non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), alcohol-related cirrhosis, and liver fibrosis in those infected with viral hepatitis (Buch et al., 2015; Mancina et al., 2016; Luukkonen et al., 2016; Thabet et al., 2016; Viitasalo et al., 2016; Krawczyk et al., 2017; Thabet et al., 2017). Based on hepatic expression quantitative trait loci analysis, it has been suggested that MBOAT7 loss of function promotes liver disease progression (Buch et al., 2015; Mancina et al., 2016; Luukkonen et al., 2016; Thabet et al., 2016; Viitasalo et al., 2016; Krawczyk et al., 2017; Thabet et al., 2017), but this has never been formally tested. Here we show that Mboat7 loss, but not Tmc4, in mice is sufficient to promote the progression of NAFLD in the setting of high fat diet. Mboat7 loss of function is associated with accumulation of its substrate lysophosphatidylinositol (LPI) lipids, and direct administration of LPI promotes hepatic inflammatory and fibrotic transcriptional changes in an Mboat7-dependent manner. These studies reveal a novel role for MBOAT7-driven acylation of LPI lipids in suppressing the progression of NAFLD.

ApoC-III Glycoforms Are Differentially Cleared by Hepatic TRL (Triglyceride-Rich Lipoprotein) Receptors.

OBJECTIVE: ApoC-III (apolipoprotein C-III) glycosylation can predict cardiovascular disease risk. Higher abundance of disialylated (apoC-III2) over monosialylated (apoC-III1) glycoforms is associated with lower plasma triglyceride levels. Yet, it remains unclear whether apoC-III glycosylation impacts TRL (triglyceride-rich lipoprotein) clearance and whether apoC-III antisense therapy (volanesorsen) affects distribution of apoC-III glycoforms. Approach and Results: To measure the abundance of human apoC-III glycoforms in plasma over time, human TRLs were injected into wild-type mice and mice lacking hepatic TRL clearance receptors, namely HSPGs (heparan sulfate proteoglycans) or both LDLR (low-density lipoprotein receptor) and LRP1 (LDLR-related protein 1). ApoC-III was more rapidly cleared in the absence of HSPG (t1/2=25.4 minutes) than in wild-type animals (t1/2=55.1 minutes). In contrast, deficiency of LDLR and LRP1 (t1/2=56.1 minutes) did not affect clearance of apoC-III. After injection, a significant increase in the relative abundance of apoC-III2 was observed in HSPG-deficient mice, whereas the opposite was observed in mice lacking LDLR and LRP1. In patients, abundance of plasma apoC-III glycoforms was assessed after placebo or volanesorsen administration. Volanesorsen treatment correlated with a statistically significant 1.4-fold increase in the relative abundance of apoC-III2 and a 15% decrease in that of apoC-III1. The decrease in relative apoC-III1 abundance was strongly correlated with decreased plasma triglyceride levels in patients. CONCLUSIONS: Our results indicate that HSPGs preferentially clear apoC-III2. In contrast, apoC-III1 is more effectively cleared by LDLR/LRP1. Clinically, the increase in the apoC-III2/apoC-III1 ratio on antisense lowering of apoC-III might reflect faster clearance of apoC-III1 because this metabolic shift associates with improved triglyceride levels.

Increased apolipoprotein C3 drives cardiovascular risk in type 1 diabetes.

Type 1 diabetes mellitus (T1DM) increases the risk of atherosclerotic cardiovascular disease (CVD) in humans by poorly understood mechanisms. Using mouse models of T1DM-accelerated atherosclerosis, we found that relative insulin deficiency rather than hyperglycemia elevated levels of apolipoprotein C3 (APOC3), an apolipoprotein that prevents clearance of triglyceride-rich lipoproteins (TRLs) and their remnants. We then showed that serum APOC3 levels predict incident CVD events in subjects with T1DM in the Coronary Artery Calcification in Type 1 Diabetes (CACTI) study. To explore underlying mechanisms, we investigated the impact of Apoc3 antisense oligonucleotides (ASOs) on lipoprotein metabolism and atherosclerosis in a mouse model of T1DM. Apoc3 ASO treatment abolished the increased hepatic Apoc3 expression in diabetic mice - resulting in lower levels of TRLs - without improving glycemic control. APOC3 suppression also prevented arterial accumulation of APOC3-containing lipoprotein particles, macrophage foam cell formation, and the accelerated atherosclerosis in diabetic mice. Our observations demonstrate that relative insulin deficiency increases APOC3 and that this results in elevated levels of TRLs and accelerated atherosclerosis in a mouse model of T1DM. Because serum levels of APOC3 predicted incident CVD events in the CACTI study, inhibiting APOC3 might reduce CVD risk in T1DM patients.

Supports: A Key Factor in Faculty Implementation of Evidence-Based Teaching.

Evidence-based teaching (EBT), such as active learning and formative assessment, benefits student learning but is not present in many college science classrooms. The choices faculty make about how to teach their science courses are influenced by their personal beliefs and motivations, as well as their departmental structures and institutional cultures. With data from 584 science, technology, engineering, and mathematics (STEM) faculty trained in EBT, we compare which of the following factors most relate to faculty's use of EBT: 1) faculty's personal motivations (e.g., teaching value, confidence, beliefs about intelligence); and 2) their experiences with their institutional teaching environments (e.g., departmental support, student enthusiasm). Faculty's perceived supports in their teaching environments (e.g., having supportive colleagues, being able to access curricular resources) were by far most predictive of their use of EBT. Faculty's personal motivations had little to no relationship when supports were included in these models. The effects were robust, even when controlling for faculty gender, minority status, and teaching experience. Much of the literature has focused on perceived barriers to EBT implementation (e.g., lack of time, constrained teaching space). The current data indicate that a focus on building supports for faculty may have the greatest impact on increasing the presence of EBT in college STEM courses.

Perceived supports and evidence-based teaching in college STEM.

BACKGROUND: Evidence-based teaching, such as active learning, is associated with increases in student learning and engagement. Although many faculty are beginning to adopt innovative practices, traditional lecture-based teaching tends to dominate college science education. What are the factors associated with faculty's decision to incorporate evidence-based teaching? While there are known barriers that limit adoption of evidence-based practices in science classrooms (e.g., lack of time, student resistance), the present work reveals that instructors' perceptions of supports (e.g., access to teaching resources, encouragement from colleagues) shows a stronger relationship to instructors' use of evidence-based teaching. RESULTS: These results come from a uniquely large dataset of college science faculty and instructors from across the USA (n = 584), who received training in evidence-based teaching. Multiple linear regression analyses of the relationship among perceived supports, barriers, and reported implementation of evidence-based practices showed that instructors report greater implementation when they perceive more social, personal, and resource supports even when barriers are also indicated as present. CONCLUSION: Faculty's perceived supports, not perceived barriers, are most strongly related to their reported implementation of evidence-based teaching. These findings suggest relevant stakeholders devote increased attention identifying and building the factors that promote evidence-based teaching in addition to reducing what inhibits it.

Hepatic monoacylglycerol acyltransferase 1 is induced by prolonged food deprivation to modulate the hepatic fasting response.

During prolonged fasting, the liver plays a central role in maintaining systemic energy homeostasis by producing glucose and ketones in processes fueled by oxidation of fatty acids liberated from adipose tissue. In mice, this is accompanied by transient hepatic accumulation of glycerolipids. We found that the hepatic expression of monoacylglycerol acyltransferase 1 (Mogat1), an enzyme with monoacylglycerol acyltransferase (MGAT) activity that produces diacyl-glycerol from monoacylglycerol, was significantly increased in the liver of fasted mice compared with mice given ad libitum access to food. Basal and fasting-induced expression of Mogat1 was markedly diminished in the liver of mice lacking the transcription factor PPARα. Suppressing Mogat1 expression in liver and adipose tissue with antisense oligonucleotides (ASOs) reduced hepatic MGAT activity and triglyceride content compared with fasted controls. Surprisingly, the expression of many other PPARα target genes and PPARα activity was also decreased in mice given Mogat1 ASOs. When mice treated with control or Mogat1 ASOs were gavaged with the PPARα ligand, WY-14643, and then fasted for 18 h, WY-14643 administration reversed the effects of Mogat1 ASOs on PPARα target gene expression and liver triglyceride content. In conclusion, Mogat1 is a fasting-induced PPARα target gene that may feed forward to regulate liver PPARα activity during food deprivation.

Angiotensinogen and Megalin Interactions Contribute to Atherosclerosis-Brief Report.

Objective- AGT (Angiotensinogen) is the unique precursor of the renin-angiotensin system that is sequentially cleaved by renin and ACE (angiotensin-converting enzyme) to produce Ang II (angiotensin II). In this study, we determined how these renin-angiotensin components interact with megalin in kidney to promote atherosclerosis. Approach and Results- AGT, renin, ACE, and megalin were present in the renal proximal convoluted tubules of wild-type mice. Hepatocyte-specific AGT deficiency abolished AGT protein accumulation in proximal tubules and diminished Ang II concentrations in kidney, while renin was increased. Megalin was most abundant in kidney and exclusively present on the apical side of proximal tubules. Inhibition of megalin by antisense oligonucleotides (ASOs) led to ablation of AGT and renin proteins in proximal tubules, while leading to striking increases of urine AGT and renin concentrations, and 70% reduction of renal Ang II concentrations. However, plasma Ang II concentrations were unaffected. To determine whether AGT and megalin interaction contributes to atherosclerosis, we used both male and female low-density lipoprotein receptor-/- mice fed a saturated fat-enriched diet and administered vehicles (PBS or control ASO) or megalin ASO. Inhibition of megalin did not affect plasma cholesterol concentrations, but profoundly reduced atherosclerotic lesion size in both male and female mice. Conclusions- These results reveal a regulatory role of megalin in the intrarenal renin-angiotensin homeostasis and atherogenesis, positing renal Ang II to be an important contributor to atherosclerosis that is mediated through AGT and megalin interactions.

Hepatocyte-Macrophage Acetoacetate Shuttle Protects against Tissue Fibrosis.

Metabolic plasticity has been linked to polarized macrophage function, but mechanisms connecting specific fuels to tissue macrophage function remain unresolved. Here we apply a stable isotope tracing, mass spectrometry-based untargeted metabolomics approach to reveal the metabolome penetrated by hepatocyte-derived glucose and ketone bodies. In both classically and alternatively polarized macrophages, [13C]acetoacetate (AcAc) labeled ∼200 chemical features, but its reduced form D-[13C]ß-hydroxybutyrate (D-ßOHB) labeled almost none. [13C]glucose labeled ∼500 features, and while unlabeled AcAc competed with only ∼15% of them, the vast majority required the mitochondrial enzyme succinyl-coenzyme A-oxoacid transferase (SCOT). AcAc carbon labeled metabolites within the cytoplasmic glycosaminoglycan pathway, which regulates tissue fibrogenesis. Accordingly, livers of mice lacking SCOT in macrophages were predisposed to accelerated fibrogenesis. Exogenous AcAc, but not D-ßOHB, ameliorated diet-induced hepatic fibrosis. These data support a hepatocyte-macrophage ketone shuttle that segregates AcAc from D-ßOHB, coordinating the fibrogenic response to hepatic injury via mitochondrial metabolism in tissue macrophages.