Nanoparticle surface-enhanced Raman spectroscopy as a noninvasive, label-free tool to monitor hematological malignancy - Supplementary Material

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Lysophosphatidic acid receptor mRNA levels in heart and white adipose tissue are associated with obesity in mice and humans

<div><p>Background</p><p>Lysophosphatidic acid (LPA) receptor signaling has been implicated in cardiovascular and obesity-related metabolic disease. However, the distribution and regulation of LPA receptors in the myocardium and adipose tissue remain unclear.</p><p>Objectives</p><p>This study aimed to characterize the mRNA expression of LPA receptors (LPA1-6) in the murine and human myocardium and adipose tissue, and its regulation in response to obesity.</p><p>Methods</p><p>LPA receptor mRNA levels were determined by qPCR in i) heart ventricles, isolated cardiomyocytes, and perigonadal adipose tissue from chow or high fat-high sucrose (HFHS)-fed male C57BL/6 mice, ii) 3T3-L1 adipocytes and HL-1 cardiomyocytes under conditions mimicking gluco/lipotoxicity, and iii) human atrial and subcutaneous adipose tissue from non-obese, pre-obese, and obese cardiac surgery patients.</p><p>Results</p><p>LPA1-6 were expressed in myocardium and white adipose tissue from mice and humans, except for LPA3, which was undetectable in murine adipocytes and human adipose tissue. Obesity was associated with increased LPA4, LPA5 and/or LPA6 levels in mice ventricles and cardiomyocytes, HL-1 cells exposed to high palmitate, and human atrial tissue. LPA4 and LPA5 mRNA levels in human atrial tissue correlated with measures of obesity. LPA5 mRNA levels were increased in HFHS-fed mice and insulin resistant adipocytes, yet were reduced in adipose tissue from obese patients. LPA4, LPA5, and LPA6 mRNA levels in human adipose tissue were negatively associated with measures of obesity and cardiac surgery outcomes. This study suggests that obesity leads to marked changes in LPA receptor expression in the murine and human heart and white adipose tissue that may alter LPA receptor signaling during obesity.</p></div

Patient characteristics and metabolic parameters.

<p>Patient characteristics and metabolic parameters.</p

Nutritional composition of research diets.

<p>Nutritional composition of research diets.</p

LPA receptor mRNA levels in myocardial tissue and cells.

<p>A) LPA1-6 mRNA levels in ventricles from chow and HFHS-fed mice in fed and 16-h fasted states (n = 5). B) LPA1-6 mRNA levels in cardiomyocytes isolated from chow and HFHS-fed mice (n = 3). C) LPA1-6 mRNA levels in HL-1 cardiomyocytes incubated in the presence or absence of 1.2 mM palmitate for 16 h (n = 3). D) LPA1-6 mRNA levels in atrial tissue from non-obese, pre-obese, and obese patients undergoing cardiac surgery (n = 6 for non-obese and pre-obese, n = 18 for obese). A-D) *<i>P</i> < 0.05, **<i>P</i> < 0.01, ***<i>P</i> < 0.001 as determined using two-way ANOVA followed by a Tukey post hoc analysis (A, D) or unpaired two-tailed t-test (B, C).</p

Experimental design and groups for LPA receptor expression analysis.

<p>Experimental layout of LPA receptor mRNA quantification in A) heart and adipose tissue from non-obese, pre-obese, and obese cardiac surgery patients, B) heart, adipose tissue, and cardiomyocytes from lean and obese mice, C) insulin sensitive and insulin resistant 3T3-L1 adipocytes, and D) insulin sensitive and insulin resistant HL-1 cardiomyocytes.</p

Unadjusted Pearson’s correlations of LPA receptors in atrial appendage from cardiac surgery patients.

<p>Unadjusted Pearson’s correlations of LPA receptors in atrial appendage from cardiac surgery patients.</p

Post-operative parameters and outcomes.

<p>Post-operative parameters and outcomes.</p

Sequence information for primers employed in qPCR analysis.

<p>Sequence information for primers employed in qPCR analysis.</p

Unadjusted Pearson’s correlations of LPA receptors in subcutaneous adipose tissue from cardiac surgery patients.

<p>Unadjusted Pearson’s correlations of LPA receptors in subcutaneous adipose tissue from cardiac surgery patients.</p