CRISPR/Cas9-mediated Angptl8 knockout suppresses plasma triglyceride concentrations and adiposity in rats

Izumi, Ryota; Kusakabe, Toru; Noguchi, Michio; Iwakura, Hiroshi; Tanaka, Tomohiro; Miyazawa, Takashi; Aotani, Daisuke; Hosoda, Kiminori; Kangawa, Kenji; Nakao, Kazuwa

doi:10.1194/jlr.m082099

Cited by 35 publications

(23 citation statements)

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“…Representative fat pads from 12-week-old male mice are shown in Figure 2A. The epididymal (Epi) fat pads from the A8 -/mice were visibly smaller and weighed significantly less than those from WT animals ( Figure 2B), which is consistent with what was seen in the A3 -/-A8 -/mice (28) and the A8 -/rat (30). The weights of the WAT-subcutaneous (SQ) fat were also lower in the A8 -/mice.…”

Section: Introductionsupporting

confidence: 79%

ANGPTL8 has both endocrine and autocrine effects on substrate utilization

Oldoni¹,

Cheng²,

Banfi³

et al. 2020

JCI Insight

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lipolysis in WAT (23, 24). Thus, A4 expression in WAT promotes flux of fatty acids toward oxidative tissues to supply them with energy substrate during fasting. A3 and A8 are functionally interdependent in vivo (11) and in vitro (25). Expression of A8 fails to suppress intravascular LPL of mice lacking A3 (11). The 2 family members form a complex in which the LPL binding domain of A8 (and not A3) is required for LPL inhibition (25). After feeding, A3 and A8 act together in the systemic circulation to inhibit LPL activity. Under these conditions, more circulating TGs bypass oxidative tissues and are hydrolyzed in WAT (7, 21). Feeding also increases expression of A8 in adipose tissue, but its role in WAT has not been defined (11, 19, 20). To begin to understand the relative roles of A8 in liver and in adipose tissue, we developed mice in which A8 is selectively inactivated in those 2 tissues. Here we show that A8 has distinct physiological functions in the 2 tissues. All detectable circulating A8 originates in the liver, and only this form of A8 complexes with A3 to inhibit intravascular lipolysis. A8 from adipose tissue makes no detectable contribution to circulating A8, but rather has local effects on substrate homeostasis. A8 expression in adipose tissue attenuates the LPL inhibitory actions of A4, thus ensuring a rapid replenishment of energy stores with feeding after a fast. Results Tissue-specific inactivation of A8. We developed C57BL/6N mice in which expression of A8 was ablated either in hepatocytes (liver-specific-A8-/mice; Ls-A8-/mice) or in adipocytes (adipose-specific-A8-/mice; As-A8-/mice). The mice were established using the Cre-lox system to remove exons 1 and 2 of A8 (Figure 1A). In the targeting construct, the neomycin gene is flanked by 2 rox sites that recombine to remove the Neo cassette, thus producing an allele with loxP sites flanking the first 2 exons of A8. Mice homozygous for the fl allele were used as controls for all experiments described in this paper, unless otherwise stated, and are referred to as WT. A8 was inactivated by crossing mice homozygous for the fl allele either with mice expressing Cre under control of the albumin promoter (B6N.Cg-Speer6-ps1 Tg(Alb-cre)21Mgn /J; Alb-Cre) (26) or with mice expressing Cre driven by the adiponectin promoter (Adipo-Cre) (27) to produce Ls-A8-/mice and As-A8-/mice, respectively. Both sexes transmitted the inactivated A8 allele, and the genotypes of the offspring conformed to the expected Mendelian ratios (Supplemental Table 1; supplemental material available online with this article; https://doi.org/10.1172/jci.insight.138777DS1). Litter sizes were similar in KO and WT mice (Supplemental Table 2). Because A8 is not expressed in the fasting state, all experiments were performed in refed conditions unless otherwise stated. The mean body weights of the C57BL/6N A8-/mice were similar to those of WT controls at 9-10 weeks of age (Figure 1B). The mean total fat mass was significantly lower in the total A8-/mice (3.0± 0.2 vs. 1.7± 0.1 g) but not in th...

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Section: Introductionsupporting

confidence: 79%

ANGPTL8 has both endocrine and autocrine effects on substrate utilization

Oldoni¹,

Cheng²,

Banfi³

et al. 2020

JCI Insight

View full text Add to dashboard Cite

show abstract

“…The relationship between ANGPTL8 and triglyceride has also been deeply excavated. A recently published paper reported that ANGPTL8 knockout mediated by CRISPR-Cas9 system led to reduced plasma triglyceride levels [ 26 ], which verified conclusions from animal experiments—ANGPTL8 was found to inhibit LPL activity and elevate plasma triglyceride partly via ANGPTL3 activation after feeding [ 14 ]. A prospective cohort study conducted in Korean children revealed that baseline ANGPTL8 was associated with future changes in triglyceride levels [ 27 ].…”

Section: Discussionsupporting

confidence: 53%

The negative effect of ANGPTL8 on HDL-mediated cholesterol efflux capacity

Luo

Zhang

Peng

et al. 2018

Cardiovasc Diabetol

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BackgroundIt is well known that angiopoietin-like protein 8 (ANGPTL8) exerts its effects on lipid metabolism through the inhibition of lipoprotein lipase and subsequent elevation of plasma triglyceride. However, it is not clear whether ANGPTL8 could affect lipid metabolism via other pathways. The study was aimed to investigate the effects of ANGPTL8 on the function of high-density lipoprotein (HDL), which plays a protective role in atherosclerosis progression.MethodsTwo hundred and ten subjects were recruited. Plasma ANGPTL8 was measured by enzyme-linked immunosorbent assays. Cholesterol efflux capacity was chosen as the biomarker of HDL function and measured via H3-cholesterol loading THP-1 cell models.ResultsANGPTL8 exhibited no significant difference between CAD group and nonCAD group, but ANGPTL8 in DM group was significantly higher than that in the nonDM group [568.3 (406.2–836.8) vs 458.2 (356.8–755.6), P = 0.023]. Compared to controls, subjects in CAD group and DM group exhibited significantly lower cholesterol efflux capacity [CAD: 14.58 ± 2.06 vs 12.51 ± 2.83%, P < 0.0001; DM: 13.62 ± 2.57 vs 12.34 ± 3.16%, P = 0.0099]. ANGPTL8 was inversely correlated with cholesterol efflux capacity (r = − 0.188, P < 0.01). Regression analysis revealed that plasma ANGPTL8 was an independent contributor to cholesterol efflux capacity (standardized β = − 0.143, P = 0.023).ConclusionANGPTL8 presents a negative effect on HDL-mediated cholesterol efflux capacity.Electronic supplementary materialThe online version of this article (10.1186/s12933-018-0785-x) contains supplementary material, which is available to authorized users.

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“…The association of ANGPTL8 with insulin resistance [18,26,27], blood glucose [28,29], C-peptide and glycosylated haemoglobin (HbA1C) have not offered clear results [30]. In animals, a positive correlation of circulating ANGPTL8 with triglyceride levels exists [10,[31][32][33]. These facts were also confirmed in humans [34].…”

Section: Introductionmentioning

confidence: 99%

Evidences for Expression and Location of ANGPTL8 in Human Adipose Tissue

Catalano-Iniesta

Robledo

Iglesias-Osma

et al. 2020

JCM

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The metabolism of triglycerides (TGs) is regulated, among others, by the lipoprotein lipase (LPL) that hydrolyses the TGs on endothelial cells. In turn, LPL is inhibited by the ANGPTLs family of proteins, such as ANGPTL3, 4, and, 8; the latter is the least known. In this work, we have tried to establish the expression and localisation of the Angiopoietin-like 8 (ANGPTL8) protein in the visceral adipose tissue (VAT) of morbid-obese and non-obese patients. 109 subjects (66 women and 43 men) undergoing laparoscopic surgery participated in this study. A blood sample and a portion of the VAT were obtained, and the patients were classified according to their Body Mass Index (BMI) as non-obese (19.5–30 kg/m2) and morbid-obese (40–50 kg/m2). No significant changes in ANGPTL8 plasma levels were determined by EIA in obese patients. The immunocytochemistry and Western blotting showed the presence of increased ANGPTL8 in morbid-obese patients (p < 0.05). In-situ hybridisation and a real time polymerase chain reaction (RT-PCR) confirmed that the mRNA that encodes ANGPTL8 was present in adipocytes, without differences in their nutritional state (p = 0.89), and even in the endothelial cells. Our data suggests that ANGPT8 plasmatic levels do not change significantly in patients with morbid obesity, although there is a modest difference related to gender. Besides, we demonstrate that in visceral adipose tissue, ANGPTL8 is well defined in the cytoplasm of adipocytes coexisting with perilipin-1 and its mRNA, also is present in endothelial cells. These findings suggest the possibility that among other functions, ANGPTL8 could perform either a paracrine and/or an endocrine role in the adipose tissue.

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CRISPR/Cas9-mediated Angptl8 knockout suppresses plasma triglyceride concentrations and adiposity in rats

Cited by 35 publications

References 50 publications

ANGPTL8 has both endocrine and autocrine effects on substrate utilization

ANGPTL8 has both endocrine and autocrine effects on substrate utilization

The negative effect of ANGPTL8 on HDL-mediated cholesterol efflux capacity

Evidences for Expression and Location of ANGPTL8 in Human Adipose Tissue

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