Citations

Introduction: Plasma aldosterone (Aldo) increases Na and water reabsorption and mediates inflammation. Our recent findings in a general population cohort showed Aldo strongly associated with hypertension (HTN), obesity, chronic kidney disease (CKD) and lower natriuretic peptide levels (NPs) during a first visit (V1), even when Aldo was within the normal range. It is undefined whether Aldo itself (V1) and/or its increase after 4-year (second visit, V2) can predict new onset cardiovascular and metabolic diseases. Hypotheses: We hypothesized that: 1) Aldo-V1 predicts new onset disease at V2; 2) The increase of Aldo between V1 and V2 predicts new onset disease-V2; and 3) Aldo-V2 remains associated with cardiovascular and metabolic disease-V2. Methods: We defined the correlations between Aldo-V1 and Aldo-V2 with cardiometabolic diseases and medications at V2, in a general community cohort (N=1368) from Olmsted County, MN. Associations were adjusted for age, sex and BMI; Aldo was analyzed as continuous variable and according to tertiles. Results: After 4 years, Aldo-V1 continuous variable predicted new onset of central obesity (p=0.0113, OR=1.36, CI=1.07-1.73), HTN (p=0.0012, OR=1.36, CI=1.13-1.63) and use of antilipemic drugs (p=0.0119, OR=1.25, CI=1.05-1.48); while Aldo-V1 in the 3 rd tertile predicted new onset of type 2 diabetes (T2DM) (p=0.0392, OR=1.96, CI=1.03-3.70), use of antilipemic therapy (p=0.0162, OR=1.59, CI=1.09-2.31) and HTN (p=0.049, OR1.44, CI=1.00-2.08) at V2. Secondly, we observed that the increase in Aldo between V1 and V2 predicted new HTN-V2. Finally, we found higher Aldo-V2, but still within the normal range and Aldo-V2 remained associated with HTN, obesity and CKD, when analyzed as continuous variable and when in the 3 rd tertile. Conclusions: In the general community, Aldo-V1 predicts new onset HTN, central obesity, T2DM and use of antilipemic therapy. Importantly, the increase of aldosterone between V1 and V2 also predicts new onset HTN. Finally, Aldo-V2 correlates with HTN, obesity and CKD as we found at V1. Further studies are warranted to evaluate aldosterone as therapeutic target to delay disease progression, and to support its role as prognostic biomarker to identify high-risk subjects.

Abstract Background: Engineered toxin bodies (ETBs), composed of an engineered Shiga-like Toxin A subunit genetically fused to an antibody-binding domain, can force receptor internalization, induce potent cell-kill via enzymatic and permanent inactivation of ribosomes, and may not be subject to resistance mechanisms of other targeted agents. MT-5111, a de-immunized 55 kD ETB targeting HER2 in solid tumors, also binds to an epitope distinct from trastuzumab and pertuzumab, which may permit combination strategies with other HER2 targeting agents. Methods: The primary objective is to determine maximum tolerated dose (MTD) of MT-5111 monotherapy in adult patients (pts) with advanced HER2+ solid tumors. Secondary objectives are PK, efficacy, and immunogenicity. Using a modified 3+3 design, the dose-escalation part of the study includes the following 7 cohorts: 0.5, 1, 2, 3, 4.5, 6.75, and 10 µg/kg. Three dose-expansion cohorts will follow for HER2+ breast cancer, gastro-esophageal cancer, and any other HER2+ tumors. All pts will receive MT-5111 weekly as a 30-min IV infusion in each 21-d treatment (tx) cycle (C) until disease progression (PD), unacceptable toxicity, death, or withdrawn consent (NCT04029922). Results: As of the data cut in December 2020, 16 pts were treated; cancer types included breast (n=6), gastric (n=1), colon (n=1), gallbladder (n=5), and other solid tumors (n=3). Mean age was 64 years (range, 34-78); 37.5% were male. Pts received a median of 4 prior lines of systemic therapies (range, 1-8). No G4 or G5 TEAEs occurred. Six pts had 11 G3 TEAEs; the most common were increased AST and dyspnea (both n=2). Three pts had tx-emergent serious adverse events (abdominal distension [n=1]; dyspnea [n=2]). Tx-related TEAEs occurred in 8 (50%) pts; the most common was fatigue (n=5, 31.3%) and all were ≤ grade 2 in nature, except for one grade 3 event of dyspnea. No cardiac TEAEs, clinically significant changes in cardiac biomarkers (troponin, electrocardiogram, left ventricular ejection fraction), or cases of capillary leak syndrome were observed. Fifteen pts discontinued with PD; 1 pt in cohort 5 (4.5 µg/kg) is on tx with stable disease. To date, no DLTs have been observed and the MTD has not been reached. One pt in cohort 2 (1 µg/kg) had resolution of all hepatic lesions (sub-centimeter lesions pre-tx) at the end of C8; however, the pt came off study due to clinical progression at the end of C10. PK data for the first 5 cohorts matched simulations based on non-human primate studies. Conclusions: MT-5111 was well tolerated with no clinically significant cardiotoxicity. Continued dose escalations are ongoing. Citation Format: Zev A. Wainberg, Monica M. Mita, Minal A. Barve, Erika P. Hamilton, Andrew J. Brenner, Frances Valdes, Daniel Ahn, Joleen Hubbard, Jason Starr, Christine Burnett, Joshua Pelham, Eric T. Williams, Banmeet S. Anand, Thomas Strack, Andrés Machado Sandri, Brian A. Van Tine. Phase 1 study of the novel immunotoxin MT-5111 in patients with HER-2+tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr CT130.

Background CD‐NP is a novel chimeric natriuretic peptide (NP) consisting of the full‐length human C‐type natriuretic peptide (CNP), a NP receptor (NPR)‐B agonist, and the C‐terminus of Dendroaspis NP (DNP), a NPR‐A agonist. In the first‐in‐human clinical trial, CD‐NP activated plasma and urinary cGMP, the second messenger for the NPs, and increased Na + excretion in normal subjects. Here we report for the first time plasma aldosterone (ALDO) and angiotensin II (ANG II) levels. Methods In the double‐blind, PLB‐controlled phase of this trial, adults received CD‐NP 17.5 ng/kg/min (n = 6) or PLB (n = 4) i.v. for 4 hours. Plasma ALDO and ANG II were quantified by RIA. Data were analyzed by 2‐tailed paired t‐test and signed‐rank (S‐R) test (SAS v9). Results In subjects who received CD‐NP, plasma ALDO (mean±SD) decreased from a baseline of 21.9±2.7 to 9.5±3.2 ng/dL (P<0.001 t‐test, P<0.05 S‐R test) at the end of infusion. Plasma ANG II tended to decrease (29.2±5.2 to 23.0±3.5 ng/L, P=0.086 t‐test, P=0.69 S‐R test). In the PLB group, ALDO (20.3±4.2 to 13.6±4.5 ng/dL) and ANG II (23.3±2.2 to 19.8±8.2 ng/L) did not change. Conclusion CD‐NP suppressed plasma ALDO in normal subjects, consistent with NPR‐A activation by a novel NP with dual NPR‐A and NPR‐B activating properties. CD‐NP may be of therapeutic potential in disease states that are characterized by ALDO activation, such as in heart failure warranting further studies. Sources of funding: Mayo Foundation and Nile Therapeutics, Inc.

Decreased renal function is associated with increased CV morbidity and mortality. The changes mild renal impairment may cause structurally/molecularly in the heart remain unclear. We hypothesized that mild renal dysfunction produced by unilateral nephrectomy(UNX)results in changes in ventricular structure and gene expression in the absence of hypertension or vol overload. Wistar rats were used[sham (S; n=10) and UNX; n=9]. At 4 weeks, GFR was determined by inulin clearance and RBF by PAH. Hearts and kidneys were harvested for histological analysis. Cardiac function assessed by echo. Genomewide microarray analysis was performed on LV myocardium (Affymetrix). Glomerular hypertrophy was observed in UNX. GFR tended to decrease with a reduction in RBF (S:8±1, Nx:5±1 ml/min, p<0.005). Blood pressure (BP), Na and water excretions were maintained without activation of PRA, aldo or BNP. LVEF and LV diameters were normal. Microarray analysis of LV revealed that 278 genes changed with UNX (1.5 fold, P<0.05) related to cell growth, bone remodeling and muscle contraction (Z value>2). Importantly, Pic Red staining in the LV of UNX revealed greater fibrosis (S:2±0.1, Nx:4±0.4 %, p<0.001). We conclude that mild renal insufficiency initiates myocardial gene responses and fibrosis independent of alterations in the circulating RAAS, BNP, BP, vol overload or systolic dysfunction.

Studies in human myocardial infarction (MI) suggest that even in the absence of heart failure (HF) alterations in renal function may occur and contribute to poor outcomes. We hypothesized that even without preexisting renal disease, renal functional, structural and molecular changes would be present following MI. Wistar rats were used, Sham(S;n=10) and MI(n=9). 3 weeks after MI, GFR was determined by inulin clearance. Blood was obtained for PRA and aldosterone. Cardiac function was assessed by echo. Genomewide microarray analysis was performed on kidney cortex (KC) and medulla (KM) (Affymetrix). EF decreased after MI (S:63±2, MI:43±6%, p<0.01) and LVEDd increased. PRA, aldosterone and blood pressure (BP) did not change. There was no HF as Na and water excretion was maintained. GFR tended to decrease. Pic Red staining in the KC and KM after MI showed greater fibrosis (KC S:1±0.2, MI:3±0.6%, p<0.001 and KM S:1±0.2, MI:19±6%, p<0.005). Microarray analysis showed that 303 genes changed in KM and 407 genes in KC after MI (1.5 fold, P<0.05) related to structure morphogenesis and metabolic processes(Z value>2). We conclude that experimental MI results in renal structural remodeling characterized by cortical and medullary fibrosis with a mild reduction in GFR and extensive modulation of molecular pathways related to renal growth and morphogenesis underscoring a heart‐kidney connection after MI.

Background CBB‐NP is a novel fully humanized designer natriuretic peptide (NP) integrating the 17‐amino‐acid ring of human C‐type NP (CNP) with the N‐ and the C‐termini of human B‐type NP (BNP). CBB‐NP exerted natriuretic, diuretic, renal‐preserving and cardiac‐unloading actions in anesthetized dogs. In this investigation, we compared for the first time the cGMP stimulating actions of CBB‐NP vs human atrial NP (ANP), BNP and CNP in human aortic endothelial cells (HAEC). Methods CBB‐NP was synthesized (SPKMVQGSGCFGLKLDRIGSMSGLGCKVLRRH with a disulfide bond joining the C residues). HAEC were incubated with ANP, BNP, CNP or CBB‐NP for 10 min (10 −6 M). Cyclic GMP was quantified by RIA. Results CBB‐NP and CNP increased cGMP (pmol/mL) vs control (0.108±0.020 and 0.174±0.036 vs 0.001±0.001, P<0.01 and <0.001, respectively). CBB‐NP elicited a greater cGMP response vs ANP (0.027±0.002, P<0.05), whereas comparable responses were observed vs BNP and CNP. When the native NPs were compared, a greater cGMP stimulating response was detected with CNP vs ANP and BNP (P<0.001 and <0.01, respectively). Conclusions The novel designer NP, CBB‐NP, stimulates cGMP in human aortic endothelial cells to a comparable extent as compared to the native BNP and CNP. Moreover, its cGMP stimulating action is greater than that of ANP. The therapeutic potential of CBB‐NP in models of cardiovascular diseases warrants further investigation. Supported by NIH, HFSA and ASCPT

Background The 17‐amino‐acid (AA) ring of C‐type natriuretic peptide (CNP) is essential for activation of NP receptor (NPR)‐B and the 2 nd messenger cGMP. CNP induces minimal hypotension as NPR‐B is more abundantly found in veins vs arteries, but possesses limited renal properties. We designed a novel peptide fusing the CNP ring with AAs from ANP and BNP to test our hypothesis that this unique peptide, CAB‐NP, would exert enhanced renal actions without inducing excessive hypotension. Methods CAB‐NP was synthesized (SLRRSSCFGLKLDRIGSMSGLGCKVLRRH) and infused (21 pmol/kg/min iv) in 5 anesthetized dogs. GFR was assessed by inulin clearance. Mean±SE, P<.05*, <.01 † , BL vs 30 min infusion. Results CAB‐NP increased plasma cGMP (3.5±0.6 to 10.9±1 † pmol/mL), urinary cGMP excretion (980±96 to 4587±825* pmol/min), net renal cGMP generation (823±88 to 4091±824* pmol/min), urine flow (0.2±.03 to 1.2±.4* mL/min) and Na + excretion (19±7 to 191±55* †Eq/min). Proximal (85±3 to 68±1 † %) and distal (98±1 to 91±2 † %) FRNa were reduced; GFR was preserved (46±4 to 45±3 mL/min). MAP mildly decreased (121±4 to 111±5* mmHg). PCWP (6±1 to 4±1 † mmHg) and RAP (3±1 to 2±1* mmHg) were reduced. Plasma renin activity (11±1 to 2±0.3 † ng/mL/hr) and aldosterone (15±4 to 7±2* ng/dL) were suppressed. Conclusions CAB‐NP exerts favorable in vivo actions without excessively lowering BP. The therapeutic potential of CAB‐NP warrants further studies. Supported by the NIH, HFSA, and ASCPT.

Decreased renal function is associated with increased CV morbidity and mortality. However, the impact of mild renal dysfunction upon the heart remains to be fully elucidated. It is thought that a mild reduction in renal mass is not deleterious to the heart, yet exploration of the impact of unilateral nephrectomy (Nx) on myocardial gene responses is undefined. We hypothesized that even when ventricular function and structure are grossly preserved after Nx, such a reduction in renal mass and function would result in significant alterations in the cardiac genome. Wistar rats, Sham(S)(n=10), Nx(n=9). 4 weeks after Nx we assessed proteinuria, UNaV, UcGMP and BNP. Cardiac function was assessed by echo. GFR was determined by inulin clearance and renal blood flow (RBF) by PAH. Blood was obtained for BNP, aldosterone and cGMP. Kidneys and hearts were harvested. Gene microarrays were conducted of left ventricle (LV) myocardium (Affymetrix GeneChip® Rat Genome 230 2.0). GFR strongly tended to decrease and RBF was decreased in Nx (S:8±1, Nx:5±1 ml/min, p<0.005). Glomerular volume was increased in Nx (S:1±0.04, Nx:1.6±0.1 u 3 ×10 6 , p<0.001) while UNaV and volume were not different. There was no proteinuria in Nx. EF, LV end systolic and diastolic diameters were normal. Blood pressure (BP) was unchanged. There was no activation of the systemic RAAS or BNP. Importantly, 278 cardiac genes changed their expression after Nx (1.5 fold, P<0.05). 11% were increased and 89% decreased. Pathways involved included cell adhesion, growth and proliferation. We conclude that Nx, independently of alterations in circulating RAAS or BNP, BP or myocardial structure and function, results in widespread changes in myocardial gene expression. Further long‐term studies are needed to define the impact of these early gene responses to myocardial structure and function.

Introduction: Atrial natriuretic peptide (ANP) plays an important role in blood pressure (BP) regulation via the second messenger cGMP. ANP also induces lipolysis by increasing plasma levels of non-esterified fatty acids (NEFA). MANP is a new Mayo Clinic designed ANP analog consisting of the native ANP with a novel 12-amino-acid C-terminus extension. In experimental models, MANP is more potent than ANP in reducing BP and inducing natriuresis. These actions are mediated by the activation of the pGC-A receptor and generation of cGMP. To date, the BP pressure lowering, cGMP generating and lipolytic actions of MANP remain unexplored along with its safety and tolerability in subjects with hypertension (HTN) associated with metabolic disease. Hypothesis: In subjects with HTN, obesity and metabolic syndrome (MS), MANP administration is safe, well tolerated, activates cGMP and exerts BP lowering and lipolytic properties. Methods: Double-blind, placebo-control clinical trial in 22 subjects (17 receiving MANP) with HTN, obesity and MS involving a single subcutaneous (SQ) injection of MANP (2.5 μg/Kg) or placebo. Subjects were monitored and cardiovascular, cGMP and metabolic parameters were collected during 24 hours after the injection. Results: In all subjects, MANP was safe and well-tolerated with no serious adverse side effects. MANP increased mean cGMP plasma levels after 30 minutes (delta: 4.8 ± 2.0 pmol/mL, p=0.02) and 1 hour (delta: 2.9 ±1.3 pmol/mL, p=0.03) when compared to baseline. Six hours after MANP injection systolic BP decreased by 5.7 ± 2.9 mmHg (p=0.06) whereas subjects receiving placebo had no decrease in blood pressure values compared to baseline. During fasting period, one hour after the administration of MANP, NEFA plasma levels increased by 108.2 ± 37.4 μM (p=0.01). Conclusions: In subjects with HTN, obesity and MS, SQ MANP is safe, well tolerated and activates the pGC-A receptor as measured by a significant increase in the second messenger cGMP. Subjects receiving MANP showed a borderline significant decrease in systolic BP and a significant increase in NEFA circulating levels supporting anti-hypertensive and lipolytic properties. Our findings support MANP as a potential novel therapy for HTN associated with metabolic disease.