MicroRNA‐33 Deficiency Reduces the Progression of Atherosclerotic Plaque in ApoE −/− Mice
Open accessAbstract
Background: Cholesterol efflux from cells to apolipoprotein A‐I (apoA‐I) acceptors via the ATP ‐binding cassette transporters ABCA 1 and ABCG 1 is thought to be central in the antiatherogenic mechanism. Micro RNA (miR)‐33 is known to target ABCA 1 and ABCG 1 in vivo. Methods and Results: We assessed the impact of the genetic loss of miR‐33 in a mouse model of atherosclerosis. MiR‐33 and apoE double‐knockout mice (miR‐33 −/− Apoe −/− ) showed an increase in circulating HDL‐C levels with enhanced cholesterol efflux capacity compared with miR‐33 +/+ Apoe −/− mice. Peritoneal macrophages from miR‐33 −/− Apoe −/− mice showed enhanced cholesterol efflux to apoA‐I and HDL‐C compared with miR‐33 +/+ Apoe −/− macrophages. Consistent with these results, miR‐33 −/− Apoe −/− mice showed reductions in plaque size and lipid content. To elucidate the roles of miR‐33 in blood cells, bone marrow transplantation was performed in these mice. Mice transplanted with miR‐33 −/− Apoe −/− bone marrow showed a significant reduction in lipid content in atherosclerotic plaque compared with mice transplanted with miR‐33 +/+ Apoe −/− bone marrow, without an elevation of HDL‐C . Some of the validated targets of miR‐33 such as RIP 140 ( NRIP 1) and CROT were upregulated in miR‐33 −/− Apoe −/− mice compared with miR‐33 +/+ Apoe −/− mice, whereas CPT 1a and AMPK α were not. Conclusions: These data demonstrate that miR‐33 deficiency serves to raise HDL‐C , increase cholesterol efflux from macrophages via ABCA 1 and ABCG 1, and prevent the progression of atherosclerosis. Many genes are altered in miR‐33‐deficient mice, and detailed experiments are required to establish miR‐33 targeting therapy in humans.
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Written from the paper’s full text.
TL;DR
Genetic loss of miR-33 in ApoE−/− mice raises circulating HDL-C, enhances macrophage cholesterol efflux via ABCA1/ABCG1, and reduces atherosclerotic plaque size and lipid content. Bone marrow transplantation shows that loss of leukocyte miR-33 reduces plaque lipid content without raising HDL-C, implicating macrophage miR-33 in atherosclerosis progression.
Key findings
- miR-33−/−Apoe−/− mice had reduced atherosclerosis versus miR-33+/+Apoe−/− controls: en face aortic lesion area in males was 17.2±1.1% versus 24.7±1.4% (P<0.001), and proximal aortic plaque area was reduced in males (0.36±0.021 versus 0.44±0.025 mm², P=0.0314) and females (0.54±0.036 versus 0.66±0.042 mm², P=0.0372).
- HDL-C was significantly elevated in miR-33−/−Apoe−/− mice by HPLC: male HDL 33.21±2.51 versus 22.79±1.28 mg/dL and female HDL 22.71±2.52 versus 17.62±0.73 mg/dL; apoB-depleted serum from these mice promoted greater cholesterol efflux from J774 macrophages.
- Peritoneal macrophages from miR-33−/−Apoe−/− mice showed increased ABCA1/ABCG1 expression and enhanced cholesterol efflux to apoA-I and HDL-C; they were also resistant to free-cholesterol-induced apoptosis.
- Bone marrow transplantation with miR-33−/−Apoe−/− marrow reduced plaque lipid content (1.9±0.2% versus 3.0±0.4%, P=0.020) without elevating HDL-C; plaque size reduction was not statistically significant (P=0.12). RIP140 and CROT were upregulated in miR-33−/−Apoe−/− mice, whereas CPT1a and AMPKα were not.
Methods
- miR-33−/− mice were crossed with Apoe−/− mice on a C57BL/6 background; miR-33+/+Apoe−/− littermates served as controls. Mice were fed normal chow until 6 weeks, then a Western-type diet containing 0.15% cholesterol and 20% fat for 16 weeks, with atherosclerosis analyzed at 22 weeks.
- Analyses included serum lipid profiling by standard methods and HPLC, cholesterol efflux assays using J774 cells and thioglycollate-elicited peritoneal macrophages, qRT-PCR, Western blotting, flow cytometry for monocyte subsets, immunohistochemistry, en face and cross-sectional aortic lesion quantification, and bone marrow transplantation after 2×6 Gy irradiation with 5×10⁶ BM cells.
- Data are mean±SE; comparisons used Mann–Whitney U tests for n<10, unpaired two-tailed Student t tests otherwise, and one-way ANOVA with Bonferroni post hoc test for >2 groups; significance was P<0.05.
Limitations
- The paper states that many genes are altered in miR-33-deficient mice and that detailed experiments are required before miR-33 targeting therapy can be established in humans; it also notes that the macrophage effect is not a simple M1-to-M2 shift.
- The authors state that they do not know why HDL-C levels were almost the same in bone marrow transplant recipients and suggest radiation to liver and intestine may have reduced the effect of miR-33 deficiency on HDL-C; they also note that static HDL-C measurement has inherent limitations and that HDL-C function in Apoe−/− mice may be altered because apoE is absent.
- Evident from the text: findings are in a mouse atherosclerosis model, BMT showed only a nonsignificant trend for plaque size reduction, and the authors caution that antagomir-based results may differ from genetic deletion and that longer-term safety assessment is needed.
Model-written summary. Check it against the paper before relying on it.
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Cites
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Claims the paper makes
20 of 34 cited sources checked · 8 supported · 0 flagged · accuracy not yet measured
- Premisecitation
MicroRNA-33 targets ABCA1 and ABCG1 in vivo.
“MicroRNA (miR)-33 is known to target ABCA1 and ABCG1 in vivo.”
Quote in the paperSubstantial claim2 of 4 sources checked- cites [16] Rayner KJ Suarez Y Davalos A Parathath S Fitzgerald ML Tamehiro N Fisher EA Moore KJ Fernandez-Hernando C MiR-33 contributes to the regulation of cholesterol homeostasis. Science. 2010; 328: 1570- 1573 20466885 PMC3114628 source
In the in vivo mouse experiment (passage [6], labeled Abstract), miR-33 significantly reduced hepatic ABCA1 expression, but the decrease in ABCG1 protein was not statistically significant. The paper also identifies ABCG1 as a miR-33 target only in mice (passage [2]), so it does not establish the unqualified claim that miR-33 targets both genes in vivo. (LLM, conf=0.90)
Experimental — accuracy not yet measured - cites [17] Najafi-Shoushtari SH Kristo F Li Y Shioda T Cohen DE Gerszten RE Naar AM MicroRNA-33 and the SREBP host genes cooperate to control cholesterol homeostasis. Science. 2010; 328: 1566- 1569 20466882 PMC3840500 source
The Results show that miR-33 targets the ABCA1 3′UTR, and the mouse experiment reports an HDL increase consistent with miR-33a regulation of ABCA1-dependent cholesterol efflux in vivo. The passages do not show that miR-33 targets ABCG1 in vivo; a search for ABCG1 returned no passage addressing it. (LLM, conf=0.87)
Experimental — accuracy not yet measured - cites [18] Marquart TJ Allen RM Ory DS Baldan A MiR-33 links SREBP-2 induction to repression of sterol transporters. Proc Natl Acad Sci USA. 2010; 107: 12228- 12232 20566875 PMC2901433 source
- cites [22] Horie T Ono K Horiguchi M Nishi H Nakamura T Nagao K Kinoshita M Kuwabara Y Marusawa H Iwanaga Y Hasegawa K Yokode M Kimura T Kita T MicroRNA-33 encoded by an intron of sterol regulatory element-binding protein 2 (SREBP2) regulates HDL in vivo. Proc Natl Acad Sci USA. 2010; 107: 17321- 17326 20855588 PMC2951399 source
- Premisecitation
miR-33 controls cholesterol homeostasis based on knockdown experiments using antisense technology.
“Recent reports, including ours, have indicated that miR-33 controls cholesterol homeostasis based on knockdown experiments using antisense technology.16–18”
Quote in the paperSubstantial claim2 of 3 sources checked- cites [16] Rayner KJ Suarez Y Davalos A Parathath S Fitzgerald ML Tamehiro N Fisher EA Moore KJ Fernandez-Hernando C MiR-33 contributes to the regulation of cholesterol homeostasis. Science. 2010; 328: 1570- 1573 20466885 PMC3114628 source
- cites [17] Najafi-Shoushtari SH Kristo F Li Y Shioda T Cohen DE Gerszten RE Naar AM MicroRNA-33 and the SREBP host genes cooperate to control cholesterol homeostasis. Science. 2010; 328: 1566- 1569 20466882 PMC3840500 source
- cites [18] Marquart TJ Allen RM Ory DS Baldan A MiR-33 links SREBP-2 induction to repression of sterol transporters. Proc Natl Acad Sci USA. 2010; 107: 12228- 12232 20566875 PMC2901433 source
- Interpretationcitation
The reduction in leukocytes observed in miR-33−/−Apoe−/− mice may have had beneficial effects on atherosclerosis.
“Because leukocytosis enhances the progression of atherosclerosis, the reduction in leukocytes observed in miR-33−/−Apoe−/− mice may have had a beneficial effects on atherosclerosis.”
Quote in the paperNot a substantial claim1 of 1 sources checked- cites [45] Coller BS Leukocytosis and ischemic vascular disease morbidity and mortality: is it time to intervene?. Arterioscler Thromb Vasc Biol. 2005; 25: 658- 670 15662026 source
- Findingown data
miR-33−/−Apoe−/− mice show reduced atherosclerotic plaque size compared with miR-33+/+Apoe−/− mice.
“Consistent with these results, miR-33−/−Apoe−/− mice showed reductions in plaque size and lipid content.”
Quote in the paperSubstantial claim - Findingown data
Peritoneal macrophages from miR-33−/−Apoe−/− mice show enhanced cholesterol efflux to apoA-I compared with miR-33+/+Apoe−/− macrophages.
“Peritoneal macrophages from miR-33−/−Apoe−/− mice showed enhanced cholesterol efflux to apoA-I and HDL-C compared with miR-33+/+Apoe−/− macrophages.”
Quote in the paperSubstantial claim
- Findingown data
Peritoneal macrophages from miR-33−/−Apoe−/− mice show enhanced cholesterol efflux to HDL-C compared with miR-33+/+Apoe−/− macrophages.
“Peritoneal macrophages from miR-33−/−Apoe−/− mice showed enhanced cholesterol efflux to apoA-I and HDL-C compared with miR-33+/+Apoe−/− macrophages.”
Quote in the paperSubstantial claim - Findingown data
RIP140 (NRIP1) and CROT are upregulated in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice.
“Some of the validated targets of miR-33 such as RIP140 (NRIP1) and CROT were upregulated in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice, whereas CPT1a and AMPKα were not.”
Quote in the paperNot a substantial claim - Findingown data
Mice transplanted with miR-33−/−Apoe−/− bone marrow do not show elevated HDL-C.
“Mice transplanted with miR-33−/−Apoe−/− bone marrow showed a significant reduction in lipid content in atherosclerotic plaque compared with mice transplanted with miR-33+/+Apoe−/− bone marrow, without an elevation of HDL-C.”
Quote in the paperSubstantial claim - Findingown data
miR-33−/−Apoe−/− mice have enhanced serum cholesterol efflux capacity compared with miR-33+/+Apoe−/− mice.
“MiR-33 and apoE double-knockout mice (miR-33−/−Apoe−/−) showed an increase in circulating HDL-C levels with enhanced cholesterol efflux capacity compared with miR-33+/+Apoe−/− mice.”
Quote in the paperSubstantial claim - Findingown data
miR-33−/−Apoe−/− mice have increased circulating HDL-C levels compared with miR-33+/+Apoe−/− mice.
“MiR-33 and apoE double-knockout mice (miR-33−/−Apoe−/−) showed an increase in circulating HDL-C levels with enhanced cholesterol efflux capacity compared with miR-33+/+Apoe−/− mice.”
Quote in the paperSubstantial claim - Findingown data
Mice transplanted with miR-33−/−Apoe−/− bone marrow show reduced lipid content in atherosclerotic plaque compared with mice transplanted with miR-33+/+Apoe−/− bone marrow.
“Mice transplanted with miR-33−/−Apoe−/− bone marrow showed a significant reduction in lipid content in atherosclerotic plaque compared with mice transplanted with miR-33+/+Apoe−/− bone marrow, without an elevation of HDL-C.”
Quote in the paperSubstantial claim - Findingown data
miR-33−/−Apoe−/− mice show reduced atherosclerotic plaque lipid content compared with miR-33+/+Apoe−/− mice.
“Consistent with these results, miR-33−/−Apoe−/− mice showed reductions in plaque size and lipid content.”
Quote in the paperSubstantial claim - Findingown data
CPT1a and AMPKα are not upregulated in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice.
“Some of the validated targets of miR-33 such as RIP140 (NRIP1) and CROT were upregulated in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice, whereas CPT1a and AMPKα were not.”
Quote in the paperNot a substantial claim - Findingown data
ICAM-1-positive area in atherosclerotic plaque in miR-33−/−Apoe−/− mice tends to be less than in miR-33+/+Apoe−/− mice (P=0.127).
“The ICAM-1-positive area in miR-33−/−Apoe−/− mice tended to be less than that in miR-33+/+Apoe−/− mice (P=0.127), shown in Figure 8C and 8D.”
Quote in the paperNot a substantial claim - Findingown data
HDL-C levels in miR-33−/−Apoe−/− mice transplanted with BM of miR-33+/+Apoe−/− or miR-33−/−Apoe−/− showed only a slight increase compared with miR-33+/+Apoe−/− mice transplanted with BM of miR-33+/+Apoe−/− or miR-33−/−Apoe−/−.
“the HDL-C levels in miR-33−/−Apoe−/− mice transplanted with BM of miR-33+/+Apoe−/− or miR-33−/−Apoe−/− (Table 3) showed only a slight increase compared with miR-33+/+Apoe−/− mice transplanted with BM of miR-33+/+Apoe−/− or miR-33−/−Apoe−/− (Table 2).”
- Also stated in: § Discussion · plain
“we did not observe a significant elevation of HDL-C level in miR-33−/−Apoe−/− mice transplanted with BM of miR-33+/+Apoe−/− or miR-33−/−Apoe−/− compared with miR-33+/+Apoe−/− mice transplanted with BM of miR-33+/+Apoe−/− or miR-33−/−Apoe−/−.”
Quote in the paperSubstantial claim - Also stated in: § Discussion · plain
- Findingown data
Total leukocyte count in miR-33−/−Apoe−/− mice is significantly less than in miR-33+/+Apoe−/− mice.
“The total leukocyte count in miR-33−/−Apoe−/− mice was significantly less than that in miR-33+/+Apoe−/− mice (Figure 6A).”
Quote in the paperNot a substantial claim - Findingown data
αSMA-positive area is significantly reduced in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice (P=0.025).
“The αSMA-positive area was also significantly reduced in miR-33−/−Apoe−/− mice (P=0.025 Figure 3C and 3D).”
Quote in the paperSubstantial claim - Findingown data
Serum HDL-C levels of miR-33+/+Apoe−/− BM recipients were the same as those in miR-33−/−Apoe−/− BM recipients.
“Serum HDL-C levels of miR-33+/+Apoe−/− BM recipients were the same as those in miR-33−/−Apoe−/− BM recipients”
Quote in the paperSubstantial claim - Findingown data
ABCA1 and ABCG1 protein expression are significantly increased in PEMs from miR-33−/−Apoe−/− mice compared with PEMs from miR-33+/+Apoe−/− mice.
“mRNA expression of ABCA1 and protein expression of ABCA1 and ABCG1 were significantly increased in PEMs from miR-33−/−Apoe−/− mice compared with PEMs from miR-33+/+Apoe−/− mice (Figure 5A and 5B).”
Quote in the paperSubstantial claim - Findingown data
ApoA-I-mediated cholesterol efflux is significantly elevated in macrophages from miR-33−/−Apoe−/− mice compared with macrophages from miR-33+/+Apoe−/− mice in a dose-dependent manner.
“Both apoA-I- and HDL-C-mediated cholesterol efflux were significantly elevated in macrophages from miR-33−/−Apoe−/− compared with macrophages from miR-33+/+Apoe−/− mice in a dose-dependent manner (Figure 5C and 5D).”
Quote in the paperSubstantial claim - Findingown data
Frequency of proinflammatory Ly6Chigh monocytes in miR-33−/−Apoe−/− mice is significantly higher than in miR-33+/+Apoe−/− mice.
“The frequency of proinflammatory Ly6Chigh monocytes in miR-33−/−Apoe−/− mice was significantly higher than that in miR-33+/+Apoe−/− mice (Figure 6B through 6E).”
Quote in the paperNot a substantial claim - Findingown data
ApoB-depleted serum from miR-33−/−Apoe−/− mice significantly promotes cholesterol efflux in J774 macrophages.
“ApoB-depleted serum from miR-33−/−Apoe−/− mice significantly promoted cholesterol efflux in J774 macrophages (Figure 4E).”
Quote in the paperSubstantial claim - Findingown data
Lesional apoptosis measured by anti-ssDNA staining is significantly reduced in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice (P=0.0493).
“There was a significant reduction in lesional apoptosis, as measured by anti-ssDNA staining (P=0.0493; Figure 3G and 3H).”
Quote in the paperNot a substantial claim - Findingown data
iNOS-positive area in atherosclerotic plaque in miR-33−/−Apoe−/− mice is significantly less than in miR-33+/+Apoe−/− mice (P=0.0057).
“As shown in Figure 7A and 7B, the iNOS-positive area in atherosclerotic plaque in miR-33−/−Apoe−/− mice was significantly less than that in miR-33+/+Apoe−/− mice (P=0.0057).”
Quote in the paperSubstantial claim - Findingown data
The ICAM-1-positive area in cross-sections of proximal aorta did not differ significantly between male miR-33−/−Apoe−/− and miR-33+/+Apoe−/− mice (P=0.127).
“Quantification of the ICAM-1-positive area in cross-sections of proximal aorta in male mice. Values are mean±SE (n=10); P=0.127.”
Quote in the paperNot a substantial claim - Findingown data
mRNA levels of M2 markers IL-10 and Chi3l3 and M1 marker IL-6 in PEMs from miR-33−/−Apoe−/− mice are significantly elevated compared with those from miR-33+/+Apoe−/− mice.
“However, our experiments indicated that the mRNA levels of both M2 markers such as IL-10 and Chi3l3 mRNA and an M1 marker such as IL-6 in PEMs from miR-33−/−Apoe−/− mice were significantly elevated compared with those from miR-33+/+Apoe−/− mice (Figure 7G).”
Quote in the paperSubstantial claim - Findingown data
Annexin V-positive cells were significantly increased in PEMs from miR-33+/+Apoe−/− mice compared with those from miR-33−/−Apoe−/− mice after free cholesterol loading.
“Annexin V–positive cells were significantly increased in PEMs from miR-33+/+Apoe−/− mice compared with those from miR-33−/−Apoe−/− mice after FC loading (Figure 11B).”
Quote in the paperSubstantial claim - Findingown data
No difference in collagen area is noted between miR-33−/−Apoe−/− and miR-33+/+Apoe−/− mice.
“However, no difference was noted in the collagen area (Figure 3E and 3F).”
Quote in the paperNot a substantial claim - Findingown data
Proximal aortic plaque area is significantly reduced in miR-33−/−Apoe−/− mice of both sexes (male P=0.0314, 0.44±0.025 versus 0.36±0.021 mm2; female P=0.0372, 0.66±0.042 versus 0.54±0.036 mm2).
“Atherosclerotic lesions were significantly reduced in miR-33−/−Apoe−/− mice of both sexes (male: P=0.0314, 0.44±0.025 versus 0.36±0.021 mm2; female: P=0.0372, 0.66±0.042 versus 0.54 ± 0.036 mm2; Figure 10D and 10E).”
Quote in the paperSubstantial claim - Findingown data
No differences were observed in CPT1a and AMPKα expressions in livers of miR-33−/−Apoe−/− and miR-33+/+Apoe−/− mice.
“No differences were observed in CPT1a and AMPKα expressions.”
Quote in the paperNot a substantial claim - Findingown data
HDL-C-mediated cholesterol efflux is significantly elevated in macrophages from miR-33−/−Apoe−/− mice compared with macrophages from miR-33+/+Apoe−/− mice in a dose-dependent manner.
“Both apoA-I- and HDL-C-mediated cholesterol efflux were significantly elevated in macrophages from miR-33−/−Apoe−/− compared with macrophages from miR-33+/+Apoe−/− mice in a dose-dependent manner (Figure 5C and 5D).”
Quote in the paperSubstantial claim - Findingown data
CD3-positive cells are significantly reduced in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice (P=0.0066).
“CD3-positive cells were significantly reduced in miR-33−/−Apoe−/− compared with miR-33+/+Apoe−/− mice (P=0.0066; Figure 3A and 3B).”
Quote in the paperSubstantial claim - Findingown data
Only the HDL-C level differs between the serum of miR-33+/+Apoe−/− and miR-33−/−Apoe−/− mice by HPLC.
“These results show that only the HDL-C level differed between the serum of miR-33+/+Apoe−/− and miR-33−/−Apoe−/− mice.”
Quote in the paperSubstantial claim - Findingown data
ABCA1 mRNA expression is significantly increased in PEMs from miR-33−/−Apoe−/− mice compared with PEMs from miR-33+/+Apoe−/− mice.
“mRNA expression of ABCA1 and protein expression of ABCA1 and ABCG1 were significantly increased in PEMs from miR-33−/−Apoe−/− mice compared with PEMs from miR-33+/+Apoe−/− mice (Figure 5A and 5B).”
Quote in the paperSubstantial claim - Findingown data
The CD68-positive area did not differ significantly between mice transplanted with miR-33−/−Apoe−/− BM and miR-33+/+Apoe−/− BM (P=0.09).
“The CD68-positive area is shown in Figure 10F and 10G (P=0.09, 19.5±2.6% versus 26.6±2.6%).”
Quote in the paperNot a substantial claim - Findingown data
VCAM-1-positive area in atherosclerotic plaque in miR-33−/−Apoe−/− mice is significantly less than in miR-33+/+Apoe−/− mice (P=0.0008).
“As shown in Figure 8A and 8B, the VCAM-1-positive area in atherosclerotic plaque in miR-33−/−Apoe−/− was significantly less than that in miR-33+/+Apoe−/− mice (P=0.0008).”
Quote in the paperSubstantial claim - Findingown data
Lipid accumulation by oil red O staining was significantly decreased in mice transplanted with miR-33−/−Apoe−/− BM compared with mice with miR-33+/+Apoe−/− BM (P=0.020).
“the quantification of lipid accumulation by oil red O staining showed a significant decrease in mice transplanted with miR-33−/−Apoe−/− BM compared with mice with miR-33+/+Apoe−/− BM (P=0.020, 3.0±0.4% versus 1.9±0.2%; Figure 10D and 10E).”
Quote in the paperSubstantial claim - Findingown data
En face analysis of the total aorta shows atherosclerotic lesions throughout the aorta are significantly reduced in miR-33−/−Apoe−/− male mice compared with miR-33+/+Apoe−/− mice (P<0.001, 24.7±1.4% versus 17.2±1.1%).
“The en face analysis of the total aorta showed that atherosclerotic lesions throughout the aorta were significantly reduced in miR-33−/−Apoe−/− mice compared with miR-33+/+ Apoe−/−mice (male: P<0.001, 24.7±1.4% versus 17.2±1.1%, Figure 1B and 1C).”
Quote in the paperSubstantial claim - Findingown data
Protein level of RIP140 in miR-33−/−Apoe−/− macrophages was significantly increased compared with miR-33+/+Apoe−/− macrophages.
“Protein level of RIP140 in miR-33−/−Apoe−/− macrophages was significantly increased compared with that in miR-33+/+Apoe−/− macrophages (Figure 9F through 9H)”
Quote in the paperSubstantial claim - Findingown data
No differences are observed in IL-6 and IL-10 immunostaining between miR-33−/−Apoe−/− and miR-33+/+Apoe−/− mice.
“On the other hand, no differences were observed in IL-6 and IL-10 immunostaining (Figure 7C through 7F).”
Quote in the paperNot a substantial claim - Findingown data
HDL-C is significantly elevated in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice of both sexes.
“HDL-C was significantly elevated in miR-33−/− Apoe−/− mice compared with miR-33+/+Apoe−/− mice of both sexes (Figure 4A).”
Quote in the paperSubstantial claim - Findingown data
No apparent changes in liver histology were observed in miR-33+/+Apoe−/− and miR-33−/−Apoe−/− mice by HE staining.
“No apparent changes in histology were observed in the livers of these mice, as shown by HE staining (Figure 9E).”
Quote in the paperNot a substantial claim - Findingown data
CD68-positive area is significantly decreased in proximal aortas of miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice (P=0.0022, 24.9±1.0% versus 20.5±0.7%).
“The CD68-positive area was also significantly decreased in the proximal aortas of miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice (P=0.0022, 24.9±1.0% versus 20.5±0.7%; Figure 2C and 2D).”
Quote in the paperSubstantial claim - Findingown data
Cleaved caspase-3 was increased in PEMs from miR-33+/+Apoe−/− mice compared with those from miR-33−/−Apoe−/− mice after free cholesterol loading.
“Cleaved caspase-3 was also increased in PEMs from miR-33+/+Apoe−/− mice compared with those from miR-33−/−Apoe−/− mice after FC loading (Figure 11C).”
Quote in the paperSubstantial claim - Findingown data
Lipid accumulation by oil red O staining is significantly decreased in miR-33−/−Apoe−/− mice compared with miR-33+/+Apoe−/− mice (P=0.034, 9.8±1.1% versus 7.1±0.5%).
“Moreover, the quantification of lipid accumulation by oil red O staining showed a significant decrease in miR-33−/−Apoe−/− compared with miR-33+/+Apoe−/− mice (P=0.034, 9.8±1.1% versus 7.1±0.5%; Figure 2A and 2B).”
Quote in the paperSubstantial claim - Findingown data
The area of lipid accumulation in miR-33−/−Apoe−/− mice transplanted with miR-33−/−Apoe−/− BM was significantly reduced compared with miR-33−/−Apoe−/− mice transplanted with miR-33+/+Apoe−/− BM (P=0.027).
“Although the area of lipid accumulation in miR-33−/−Apoe−/−mice transplanted with miR-33−/−Apoe−/− BM was significantly reduced compared with that in miR-33−/−Apoe−/−mice transplanted with miR-33+/+Apoe−/− BM (P=0.027, 3.4±0.3% versus 2.2±0.3%; Figure 12D and 12E)”
Quote in the paperSubstantial claim - Findingown data
Atherosclerotic plaque formation in miR-33−/−Apoe−/− mice transplanted with miR-33−/−Apoe−/− BM was comparable with that in miR-33−/−Apoe−/− mice transplanted with miR-33+/+Apoe−/− BM.
“Atherosclerotic plaque formation in miR-33−/− Apoe−/−mice transplanted with miR-33−/−Apoe−/− BM was comparable with that in miR-33−/− Apoe−/−mice transplanted with miR-33+/+ Apoe−/− BM (0.46±0.023 versus 0.42±0.020 mm2; Figure 12B and 12C).”
- Also stated in: § Discussion · plain
“Atherosclerotic plaque size was not significantly reduced in miR-33−/−Apoe−/− recipients compared with the miR-33+/+Apoe−/− recipients transferred with the same type of BM”
Quote in the paperSubstantial claim - Also stated in: § Discussion · plain
- Findingown data
There was no difference between miR-33−/−Apoe−/− mice transplanted with miR-33−/−Apoe−/− BM and those transplanted with miR-33+/+Apoe−/− BM in CD68-positive area (P=0.37).
“there was no difference between these mice in the CD68-positive area (P=0.37, 35.3±3.5% versus 41.2±3.3%; Figure 12F and 12G).”
Quote in the paperNot a substantial claim - Findingown data
miR-33−/−Apoe−/− mice showed an increase in HDL-C in vivo compared with miR-33+/+Apoe−/− mice.
“In the current study, miR-33−/−Apoe−/− mice showed an increase in HDL-C in vivo and a decrease in atherosclerotic plaque size and lipid content compared with miR-33+/+Apoe−/− mice.”
Quote in the paperSubstantial claim - Findingown data
Some M1 and M2 markers were significantly elevated in miR-33−/−Apoe−/− PEMs compared with miR-33+/+Apoe−/− PEMs.
“Some M1 and M2 markers were significantly elevated in miR-33−/−Apoe−/− PEMs compared with miR-33+/+Apoe−/− PEMs.”
Quote in the paperNot a substantial claim - Findingown data
Total leukocyte count in miR-33−/−Apoe−/− mice was less than that in miR-33+/+Apoe−/− mice.
“We first determined that the total leukocyte count in miR-33−/−Apoe−/− mice was less than that in miR-33+/+Apoe−/−mice.”
Quote in the paperSubstantial claim - Findingown data
Loss of miR-33 significantly increased the capacity to promote cholesterol efflux.
“We found that the loss of miR-33 significantly increased the capacity to promote cholesterol efflux”
Quote in the paperSubstantial claim - Findingown data
A higher frequency of Ly6Chigh monocytes was detected in miR-33−/−Apoe−/− mice than in miR-33+/+Apoe−/− mice.
“we also detected a higher frequency of Ly6Chigh monocytes in miR-33−/−Apoe−/− mice than in miR-33+/+Apoe−/− mice”
Quote in the paperNot a substantial claim - Findingreasoning
Apoptotic cells in the lesion area were reduced in miR-33−/−Apoe−/− mice.
“In accordance with this in vitro experiment, apoptotic cells in the lesion area were also reduced in miR-33−/−Apoe−/− mice.”
Quote in the paperSubstantial claim - Findingown data
miR-33−/−Apoe−/− macrophages promoted the removal of intracellular lipid content compared with miR-33+/+Apoe−/− macrophages.
“miR-33−/−Apoe−/− macrophages promoted the removal of intracellular lipid content compared with miR-33+/+Apoe−/− macrophages.”
Quote in the paperSubstantial claim - Findingown data
miR-33−/−Apoe−/− mice showed a decrease in atherosclerotic plaque size and lipid content compared with miR-33+/+Apoe−/− mice.
“In the current study, miR-33−/−Apoe−/− mice showed an increase in HDL-C in vivo and a decrease in atherosclerotic plaque size and lipid content compared with miR-33+/+Apoe−/− mice.”
Quote in the paperSubstantial claim - Interpretationown data
miR-33 deficiency increases macrophage cholesterol efflux via ABCA1 and ABCG1.
“These data demonstrate that miR-33 deficiency serves to raise HDL-C, increase cholesterol efflux from macrophages via ABCA1 and ABCG1, and prevent the progression of atherosclerosis.”
Quote in the paperSubstantial claim - Interpretationown data
miR-33 deficiency prevents the progression of atherosclerosis.
“These data demonstrate that miR-33 deficiency serves to raise HDL-C, increase cholesterol efflux from macrophages via ABCA1 and ABCG1, and prevent the progression of atherosclerosis.”
- Also stated in: § Discussion · boosted
“Together, these data demonstrate that miR-33 deficiency serves to raise HDL-C, improve cholesterol efflux in macrophages, and prevent the progression of atherosclerosis”
Quote in the paperSubstantial claim - Also stated in: § Discussion · boosted
- Interpretationown data
miR-33 deficiency improves macrophage cholesterol efflux by increasing macrophage ABCA1 and ABCG1 expression.
“These results indicate that miR-33 deficiency improved macrophage cholesterol efflux by increasing the expressions of macrophage ABCA1 and ABCG1.”
Quote in the paperSubstantial claim - Study design and conductown data
It was impossible to observe an effect of HDL-C elevation caused by loss of miR-33 on atherosclerosis in recipients that had the same type of blood cells.
“Therefore, it was impossible to observe an effect of HDL-C elevation caused by the loss of miR-33 on atherosclerosis in recipients that had the same type of blood cells (Figures 10 and 12).”
Quote in the paperSubstantial claim - Interpretationcitation
The effect of miR-33 deletion in macrophages is not as simple as the shift from the M1 to M2 phenotype reported previously.
“Moreover, the effect of miR-33 deletion in macrophages is not as simple as the shift from the M1 to M2 phenotype reported previously.19”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [19] Rayner KJ Sheedy FJ Esau CC Hussain FN Temel RE Parathath S van Gils JM Rayner AJ Chang AN Suarez Y Fernandez-Hernando C Fisher EA Moore KJ Antagonism of miR-33 in mice promotes reverse cholesterol transport and regression of atherosclerosis. J Clin Invest. 2011; 121: 2921- 2931 21646721 PMC3223840 source
- Interpretationown data
Loss of miR-33 in blood cells reduced the lipid content of atherosclerotic plaque.
“These results showed that loss of miR-33 in blood cells reduced the lipid content of atherosclerotic plaque.”
- Also stated in: § Discussion · boosted
“The results of the present BMT experiment revealed that deletion of macrophage miR-33 significantly reduced the lipid content in atherosclerotic plaque.”
Quote in the paperSubstantial claim - Also stated in: § Discussion · boosted
- Premisecitation
There is an inverse relationship between HDL-C concentration and cardiovascular risk.
“Thus, clinical and epidemiological studies have consistently shown that there is an inverse relationship between HDL-C concentration and cardiovascular risk.”
Quote in the paperSubstantial claim - Interpretationown data
Loss of miR-33 may affect multiple pathways in both pro- and anti-inflammatory processes.
“Overall, these results demonstrate that a loss of miR-33 may have affected multiple pathways in both pro- and anti-inflammatory processes.”
Quote in the paperSubstantial claim - Premisecitation
miR-33−/− mice show 22% to 39% higher serum HDL-C levels than wild-type mice.
“We previously reported that miR-33−/− mice showed 22% to 39% higher serum HDL-C levels than wild-type mice.22”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [22] Horie T Ono K Horiguchi M Nishi H Nakamura T Nagao K Kinoshita M Kuwabara Y Marusawa H Iwanaga Y Hasegawa K Yokode M Kimura T Kita T MicroRNA-33 encoded by an intron of sterol regulatory element-binding protein 2 (SREBP2) regulates HDL in vivo. Proc Natl Acad Sci USA. 2010; 107: 17321- 17326 20855588 PMC2951399 source
- Interpretationown data
miR-33 expression in macrophages did not contribute to serum HDL-C levels.
“These results indicated that miR-33 expression in macrophages did not contribute to serum HDL-C levels”
Quote in the paperSubstantial claim - Interpretationown data
PEMs from miR-33−/−Apoe−/− mice were significantly resistant to apoptosis compared with those from miR-33+/+Apoe−/− mice after free cholesterol loading.
“Treatment of PEMs in culture with acLDL plus ACAT inhibitor (free cholesterol loading) demonstrated that PEMs from miR-33−/−Apoe−/− mice were significantly resistant to apoptosis compared with those from miR-33+/+Apoe−/− mice.”
Quote in the paperSubstantial claim - Premisereasoning
There is still a substantial risk of CVD despite statins being widely used to lower LDL-C and apolipoprotein B-containing lipoproteins.
“However, this simply means that there is still a substantial risk of CVD despite statins being widely used to lower levels of LDL-C and apolipoprotein B–containing lipoproteins.”
Quote in the paperSubstantial claim - Interpretationreasoning
Only some of the many estimated targets of miR-33 actually have an effect in vivo through chronic complete inhibition of miRNA.
“it seems that only some of these actually have any effect in vivo through the chronic complete inhibition of miRNA.”
Quote in the paperNot a substantial claim - Interpretationreasoning
The effect of miR-33 deletion in macrophages is not as simple as a shift from the M1 to the M2 phenotype.
“In any case, the effect of miR-33 deletion in macrophages is not as simple as a shift from the M1 to the M2 phenotype, as described in a previous report.19”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [19] Rayner KJ Sheedy FJ Esau CC Hussain FN Temel RE Parathath S van Gils JM Rayner AJ Chang AN Suarez Y Fernandez-Hernando C Fisher EA Moore KJ Antagonism of miR-33 in mice promotes reverse cholesterol transport and regression of atherosclerosis. J Clin Invest. 2011; 121: 2921- 2931 21646721 PMC3223840 source
- Interpretationown data
miR-33 deficiency contributes to the reduction of plaque size in the progression of advanced atherosclerosis.
“this current study indicates that miR-33 deficiency contributes to the reduction of plaque size in the progression of advanced atherosclerosis.”
Quote in the paperSubstantial claim - Interpretationown data
miR-33 deficiency raises HDL-C.
“Together, these data demonstrate that miR-33 deficiency serves to raise HDL-C, improve cholesterol efflux in macrophages, and prevent the progression of atherosclerosis”
Quote in the paperSubstantial claim - Interpretationown data
miR-33 deficiency improves cholesterol efflux in macrophages.
“Together, these data demonstrate that miR-33 deficiency serves to raise HDL-C, improve cholesterol efflux in macrophages, and prevent the progression of atherosclerosis”
Quote in the paperSubstantial claim - Study design and conductown data
MiR-33 was barely detectable in BM cells from miR-33−/−Apoe−/− BM recipients by quantitative PCR analysis for miR-33.
“MiR-33 was barely detectable in BM cells from miR-33−/−Apoe−/− BM recipients by quantitative PCR analysis for miR-33 (data not shown).”
Quote in the paperSubstantial claim - Interpretationcitation
It may be possible to inhibit miR-33a and miR-33b pharmacologically to raise HDL-C for the treatment of dyslipidemia and atherosclerosis.
“These data suggest that it may be possible to inhibit miR-33a and miR-33b pharmacologically to raise HDL-C for the treatment of dyslipidemia and atherosclerosis.”
Quote in the paperSubstantial claim - Premisecitation
miR-33 targets ABCA1 in macrophages and the liver.
“Previously, we and others showed that miR-33 targets ABCA1 in macrophages and the liver.”
Quote in the paperSubstantial claim - Interpretationreasoning
Suppression of miR-33 in macrophages may be beneficial for the prevention of plaque rupture.
“Thus, the suppression of miR-33 in macrophages may also be beneficial for the prevention of plaque rupture.”
Quote in the paperSubstantial claim- Rests on an assumption: Apoptosis of macrophages in advanced plaques may lead to plaque rupture.
- Premiseown data
ABCG1 is another target of miR-33 in macrophages.
“ABCG1 is another target in macrophages.”
Quote in the paperNot a substantial claim - Interpretationcitation
Results obtained by antisense oligonucleotide-based medicine may be different from those obtained in miR-33-deficient mice.
“the results obtained by antisense oligonucleotide-based medicine may be different from those obtained in miR-33-deficient mice.”
Quote in the paperNot a substantial claim - Interpretationreasoning
The reduction in plaque lipid content by loss of miR-33 in blood cells may be because of improved cholesterol efflux from macrophages.
“Our results showed that loss of miR-33 in blood cells reduced the lipid content of atherosclerotic plaque, which may be because of improved cholesterol efflux from macrophages.”
Quote in the paperSubstantial claim - Study design and conductown data
Successful hematopoietic reconstitution after BMT was confirmed by PCR amplification of the whole-blood genome 4 weeks after BMT and by PCR amplification of the BM and tail genomes at the time of euthanization.
“Successful hematopoietic reconstitution after BMT was confirmed by PCR amplification of the whole-blood genome 4 weeks after BMT and by PCR amplification of the BM and tail genomes at the time of euthanization.”
Quote in the paperSubstantial claim
- Classified as probably not a substantial claim. 21
- May combine two claims. 10
- The quote only partly states this claim. 1
Classified as probably not a substantial claim.
PremisecitationCholesterol efflux from cells to apolipoprotein A-I (apoA-I) acceptors via the ATP-binding cassette transporters ABCA1 and ABCG1 is central in the antiatherogenic mechanism.
“Cholesterol efflux from cells to apolipoprotein A-I (apoA-I) acceptors via the ATP-binding cassette transporters ABCA1 and ABCG1 is thought to be central in the antiatherogenic mechanism.”
Quote in the paperSubstantial claim5 of 6 sources checked- cites [10] Aiello RJ Brees D Bourassa PA Royer L Lindsey S Coskran T Haghpassand M Francone OL Increased atherosclerosis in hyperlipidemic mice with inactivation of ABCA1 in macrophages. Arterioscler Thromb Vasc Biol. 2002; 22: 630- 637 11950702 source
The Abstract says ABCA1 promotes cholesterol efflux from cells, and the Discussion reports that macrophage-specific ABCA1 inactivation increased atherosclerosis. The copy does not establish a role for ABCG1 or support the claim that both transporters mediate efflux to apoA-I acceptors. (LLM, conf=0.88)
Experimental — accuracy not yet measured - cites [11] Joyce CW Wagner EM Basso F Amar MJ Freeman LA Shamburek RD Knapper CL Syed J Wu J Vaisman BL Fruchart-Najib J Billings EM Paigen B Remaley AT Santamarina-Fojo S Brewer HB Jr ABCA1 overexpression in the liver of LDLr-KO mice leads to accumulation of pro-atherogenic lipoproteins and enhanced atherosclerosis. J Biol Chem. 2006; 281: 33053- 33065 16928680 source
The Abstract describes ABCA1-mediated cholesterol transfer to apoA-I and calls nascent HDL formation a critical step in antiatherogenic reverse cholesterol transport. The Discussion instead associates ABCG1 with cholesterol removal by mature HDL, not apoA-I acceptors. The claim is therefore supported for ABCA1 but not for the stated acceptor of ABCG1-mediated efflux. (LLM, conf=0.88)
Experimental — accuracy not yet measured - cites [12] Singaraja RR Fievet C Castro G James ER Hennuyer N Clee SM Bissada N Choy JC Fruchart JC McManus BM Staels B Hayden MR Increased ABCA1 activity protects against atherosclerosis. J Clin Invest. 2002; 110: 35- 42 12093886 PMC151034 source
The Introduction (p. 1) says ABCA1 plays a key role in cholesterol efflux to ApoA-I and links that process to protection against atherosclerosis. Our copy does not address ABCG1, so it does not support attributing the proposed mechanism to both transporters. (LLM, conf=0.90)
Experimental — accuracy not yet measured - cites [13] van Eck M Bos IS Kaminski WE Orso E Rothe G Twisk J Bottcher A Van Amersfoort ES Christiansen-Weber TA Fung-Leung WP Van Berkel TJ Schmitz G Leukocyte ABCA1 controls susceptibility to atherosclerosis and macrophage recruitment into tissues. Proc Natl Acad Sci USA. 2002; 99: 6298- 6303 11972062 PMC122943 source
The Results and Discussion (p. 6) propose ABCA1-mediated cholesterol efflux as one possible explanation for ABCA1’s antiatherosclerotic effect. The Results and Discussion (p. 3) also describe apoA-I acquiring cholesterol from tissues through ABCA1. Our copy does not address ABCG1, so it does not support attributing the proposed mechanism to both transporters. (LLM, conf=0.88)
Experimental — accuracy not yet measured - cites [14] Brunham LR Singaraja RR Duong M Timmins JM Fievet C Bissada N Kang MH Samra A Fruchart JC McManus B Staels B Parks JS Hayden MR Tissue-specific roles of ABCA1 influence susceptibility to atherosclerosis. Arterioscler Thromb Vasc Biol. 2009; 29: 548- 554 19201688 source
- cites [15] Tang C Liu Y Kessler PS Vaughan AM Oram JF The macrophage cholesterol exporter ABCA1 functions as an anti-inflammatory receptor. J Biol Chem. 2009; 284: 32336- 32343 19783654 PMC2781648 source
The Introduction says ABCA1 exports cholesterol from cells to apoA-I, and the Discussion links ABCA1-mediated cholesterol export to cardioprotection in atherosclerotic lesions. Our copy does not establish the claim about ABCG1-mediated efflux to apoA-I, so the citation supports only the ABCA1 part. (LLM, conf=0.88)
Experimental — accuracy not yet measured
Classified as probably not a substantial claim.
PremisecitationStatins can only reduce the risk of cardiovascular events by up to 50% depending on disease status and amount of statin used, leaving a large burden of residual disease risk.
“Although the lowering of LDL cholesterol (LDL-C) with the use of statins has revolutionized the treatment of atherosclerotic cardiovascular disease, statins can only reduce the risk of cardiovascular events by up to 50% depending on the disease status and the amount of statin used, which still leaves a large burden of residual disease risk.1–3”
Quote in the paperSubstantial claim1 of 3 sources checked- cites [1] Baigent C Keech A Kearney PM Blackwell L Buck G Pollicino C Kirby A Sourjina T Peto R Collins R Simes R Efficacy and safety of cholesterol-lowering treatment: prospective meta-analysis of data from 90,056 participants in 14 randomised trials of statins. Lancet. 2005; 366: 1267- 1278 16214597 source
- cites [2] Kearney PM Blackwell L Collins R Keech A Simes J Peto R Armitage J Baigent C Efficacy of cholesterol-lowering therapy in 18,686 people with diabetes in 14 randomised trials of statins: a meta-analysis. Lancet. 2008; 371: 117- 125 18191683 source
- cites [3] Baigent C Blackwell L Emberson J Holland LE Reith C Bhala N Peto R Barnes EH Keech A Simes J Collins R Efficacy and safety of more intensive lowering of LDL cholesterol: a meta-analysis of data from 170,000 participants in 26 randomised trials. Lancet. 2010; 376: 1670- 1681 21067804 PMC2988224 source
The Discussion reports that a 2–3 mmol/L LDL reduction would lower the risk of occlusive vascular events by about 40–50%, and that more intensive therapy reduces residual risk further. It does not establish 50% as a universal maximum for statins or extend that estimate to every type of cardiovascular event. (LLM, conf=0.85)
Experimental — accuracy not yet measured
Classified as probably not a substantial claim.
PremisecitationA 1 mg/dL increment in HDL-C levels is associated with a 2% decrease in the risk of coronary heart disease in men and a 3% decrease in women.
“In fact, a 1 mg/dL (0.026 mmol/L) increment in HDL-C levels was associated with a significant decrease in the risk of coronary heart disease of 2% in men and 3% in women.6”
Quote in the paperSubstantial claim1 of 1 sources checked- cites [6] Gordon DJ Probstfield JL Garrison RJ Neaton JD Castelli WP Knoke JD Jacobs DR Jr Bangdiwala S Tyroler HA High-density lipoprotein cholesterol and cardiovascular disease. Four prospective American studies. Circulation. 1989; 79: 8- 15 2642759 source
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationABCA1 acts directly as an anti-inflammatory receptor independent of its lipid transport activities.
“Moreover, ABCA1 has been reported to act directly as an anti-inflammatory receptor independent of its lipid transport activities.15”
Quote in the paperNot a substantial claim1 of 1 sources checked- cites [15] Tang C Liu Y Kessler PS Vaughan AM Oram JF The macrophage cholesterol exporter ABCA1 functions as an anti-inflammatory receptor. J Biol Chem. 2009; 284: 32336- 32343 19783654 PMC2781648 source
Classified as probably not a substantial claim.
PremisecitationStudies in mice with Abca1 deletion or overexpression generally support that ABCA1 prevents atherosclerosis by maintaining circulating HDL-C levels and cellular cholesterol efflux.
“Studies in mice in which the Abca1 locus is either deleted or overexpressed generally have supported the hypothesis that ABCA1 significantly prevents atherosclerosis by maintaining circulating HDL-C levels and cellular cholesterol efflux.10–13”
Quote in the paperSubstantial claim4 of 4 sources checked- cites [10] Aiello RJ Brees D Bourassa PA Royer L Lindsey S Coskran T Haghpassand M Francone OL Increased atherosclerosis in hyperlipidemic mice with inactivation of ABCA1 in macrophages. Arterioscler Thromb Vasc Biol. 2002; 22: 630- 637 11950702 source
The Abstract reports that selective ABCA1 inactivation in macrophages increased atherosclerosis, and describes ABCA1 as promoting cellular cholesterol efflux. But it also reports that complete deletion did not increase atherosclerosis. The Discussion concludes that ABCA1’s antiatherogenic effect in macrophages was independent of circulating HDL levels, rather than operating by maintaining them (page numbers unavailable in our copy). (LLM, conf=0.88)
Experimental — accuracy not yet measured - cites [11] Joyce CW Wagner EM Basso F Amar MJ Freeman LA Shamburek RD Knapper CL Syed J Wu J Vaisman BL Fruchart-Najib J Billings EM Paigen B Remaley AT Santamarina-Fojo S Brewer HB Jr ABCA1 overexpression in the liver of LDLr-KO mice leads to accumulation of pro-atherogenic lipoproteins and enhanced atherosclerosis. J Biol Chem. 2006; 281: 33053- 33065 16928680 source
The Discussion says ABCA1 facilitates cellular cholesterol efflux and raises plasma HDL, and that macrophage ABCA1 expression is atheroprotective. But it describes conflicting results across mouse models: the Abstract reports no change in atherosclerosis with complete ABCA1 deficiency, while this study found increased atherosclerosis with hepatic overexpression. The claim's generalization about deletion and overexpression therefore exceeds what this paper supports. (LLM, conf=0.88)
Experimental — accuracy not yet measured - cites [12] Singaraja RR Fievet C Castro G James ER Hennuyer N Clee SM Bissada N Choy JC Fruchart JC McManus BM Staels B Hayden MR Increased ABCA1 activity protects against atherosclerosis. J Clin Invest. 2002; 110: 35- 42 12093886 PMC151034 source
- cites [13] van Eck M Bos IS Kaminski WE Orso E Rothe G Twisk J Bottcher A Van Amersfoort ES Christiansen-Weber TA Fung-Leung WP Van Berkel TJ Schmitz G Leukocyte ABCA1 controls susceptibility to atherosclerosis and macrophage recruitment into tissues. Proc Natl Acad Sci USA. 2002; 99: 6298- 6303 11972062 PMC122943 source
The Results and Discussion (pp. 2, 5) show that leukocyte ABCA1 deficiency reduces macrophage cholesterol efflux and increases atherosclerotic lesions. But the paper reports no significant change in plasma HDL cholesterol in those mice (p. 3) and describes the protective effect as independent of plasma HDL (p. 6). Thus it supports ABCA1’s antiatherosclerotic role, but not the claim that this study’s effect operates by maintaining circulating HDL-C. (LLM, conf=0.88)
Experimental — accuracy not yet measured
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationThe liver and intestine are the major sources of HDL-C.
“which is consistent with the results that the liver and intestine are the major sources of HDL-C.39,40”
Quote in the paperNot a substantial claim1 of 2 sources checked- cites [39] Timmins JM Lee JY Boudyguina E Kluckman KD Brunham LR Mulya A Gebre AK Coutinho JM Colvin PL Smith TL Hayden MR Maeda N Parks JS Targeted inactivation of hepatic ABCA1 causes profound hypoalphalipoproteinemia and kidney hypercatabolism of apoA-I. J Clin Invest. 2005; 115: 1333- 1342 15841208 PMC1074680 source
The Introduction (p. 1) says the liver and intestine are the two quantitatively important sites of apoA-I synthesis and secretion, but it does not establish that both are major sources of HDL-C. The study concludes that the liver is the single most important source of plasma HDL in chow-fed mice; it does not determine the intestine’s contribution to HDL-C. (LLM, conf=0.86)
Experimental — accuracy not yet measured - cites [40] Brunham LR Kruit JK Pape TD Parks JS Kuipers F Hayden MR Tissue-specific induction of intestinal ABCA1 expression with a liver × receptor agonist raises plasma HDL cholesterol levels. Circ Res. 2006; 99: 672- 674 16946132 source
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
PremisecitationInhibition of miR-33a/b in nonhuman primates raised plasma HDL-C and lowered VLDL triglyceride levels.
“A recent report indicated that the inhibition of miR-33a/b in nonhuman primates raised plasma HDL-C and lowered VLDL triglyceride levels.47”
Quote in the paperNot a substantial claim1 of 1 sources checked- cites [47] Rayner KJ Esau CC Hussain FN McDaniel AL Marshall SM van Gils JM Ray TD Sheedy FJ Goedeke L Liu X Khatsenko OG Kaimal V Lees CJ Fernandez-Hernando C Fisher EA Temel RE Moore KJ Inhibition of miR-33a/b in non-human primates raises plasma HDL and lowers VLDL triglycerides. Nature. 2011; 478: 404- 407 22012398 PMC3235584 source
Probably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationA reduction of 2 to 3 mmol/L LDL-C by statins could reduce the risk of CVD by 40% to 50%.
“A previous meta-analysis indicated that a reduction of 2 to 3 mmol/L LDL-C by statins could reduce the risk of CVD by 40% to 50% because there was no evidence of any threshold within the cholesterol range studied.3”
Quote in the paperNot a substantial claim1 of 1 sources checked- cites [3] Baigent C Blackwell L Emberson J Holland LE Reith C Bhala N Peto R Barnes EH Keech A Simes J Collins R Efficacy and safety of more intensive lowering of LDL cholesterol: a meta-analysis of data from 170,000 participants in 26 randomised trials. Lancet. 2010; 376: 1670- 1681 21067804 PMC2988224 source
May combine two claims.
Findingown dataAtherosclerotic plaque formation in mice transplanted with miR-33−/−Apoe−/− BM tended to be reduced compared with mice transplanted with miR-33+/+Apoe−/− BM, but the difference was not statistically significant (P=0.12).
“Atherosclerotic plaque formation in mice transplanted with miR-33−/−Apoe−/− BM tended to be reduced compared with that in mice transplanted with miR-33+/+Apoe−/− BM, but this difference was not statistically significant (P=0.12, 0.53±0.028 versus 0.46±0.033 mm2; Figure 10B and 10C).”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataThere was no difference in total cholesterol, free cholesterol, cholesterol ester, or triglyceride levels in livers of miR-33+/+Apoe−/− and miR-33−/−Apoe−/− mice.
“there was no difference in total cholesterol, free cholesterol, cholesterol ester, or triglyceride levels in the livers of miR-33+/+Apoe−/− mice and miR-33−/−Apoe−/− mice (Figure 9D).”
Quote in the paperNot a substantial claimThe quote only partly states this claim.
Findingown datamiR-33 deficiency reduced the VCAM-1-positive area in atherosclerotic plaque in male mice.
“miR-33 deficiency reduced the VCAM-1-positive area in atherosclerotic plaque.”
- Also stated in: § Discussion · plain
“miR-33 deficiency also reduced the expression of VCAM-1”
Quote in the paperSubstantial claim- Also stated in: § Discussion · plain
May combine two claims.
Findingown dataIn livers of miR-33−/−Apoe−/− mice, mRNA of CROT and protein levels of ABCA1 and CROT were significantly elevated compared with miR-33+/+Apoe−/− mice.
“mRNA of CROT and protein levels of ABCA1 and CROT in miR-33−/−Apoe−/− mice were significantly elevated compared with those in miR-33+/+Apoe−/− mice.”
- Also stated in: § Discussion · plain
“ABCA1 and CROT in the livers of miR-33−/−Apoe−/− mice were upregulated compared with those in miR-33+/+Apoe−/− mice”
Quote in the paperSubstantial claim- Also stated in: § Discussion · plain
Probably not a substantial claim: may not pass S3 (load-bearing).
Findingown dataPlaque area in female mice is greater than in male mice.
“The plaque area in females was greater than that in males.”
Quote in the paperNot a substantial claimMay combine two claims.
Findingown dataABCA1, ABCG1, and RIP140 were upregulated in miR-33−/−Apoe−/− macrophages.
“ABCA1, ABCG1, and RIP140 were also upregulated in miR-33−/−Apoe−/− macrophages.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
PremisecitationBaseline HDL-C levels are a strong predictor of 12-month morbidity and mortality from cardiovascular events in patients on statins with low LDL-C undergoing stent placement for acute coronary syndromes.
“and that HDL-C levels at baseline are a strong predictor of 12-month morbidity and mortality from cardiovascular events in patients on statins with low LDL-C levels undergoing stent placement for acute coronary syndromes.5”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [5] Wolfram RM Brewer HB Xue Z Satler LF Pichard AD Kent KM Waksman R Impact of low high-density lipoproteins on in-hospital events and one-year clinical outcomes in patients with non-ST-elevation myocardial infarction acute coronary syndrome treated with drug-eluting stent implantation. Am J Cardiol. 2006; 98: 711- 717 16950168 source
Classified as probably not a substantial claim.
PremisecitationHDL-C levels under statin treatment remain an independent predictor of the risk of cardiovascular events in patients treated with statins to lower LDL-C.
“Recent studies have shown that HDL cholesterol (HDL-C) levels under statin treatment remain an independent predictor of the risk of cardiovascular events in patients who have been treated with statins to lower LDL-C levels4”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [4] Barter P Gotto AM LaRosa JC Maroni J Szarek M Grundy SM Kastelein JJ Bittner V Fruchart JC HDL cholesterol, very low levels of LDL cholesterol, and cardiovascular events. N Engl J Med. 2007; 357: 1301- 1310 17898099 source
Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Study design and conductreasoningBoth miR-33−/− and Apoe−/− mice had a BL/6 background, so miR-33−/−Apoe−/− mice also had a BL/6 background.
“Because both knockout mice had a BL/6 background, miR-33−/−Apoe−/− mice also had a BL/6 background.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing).
PremisecitationAntisense inhibition of miR-33 results in regression of atherosclerotic plaque volume in LDL-receptor-deficient mice.
“Moreover, antisense inhibition of miR-33 resulted in a regression of the atherosclerotic plaque volume in LDL-receptor-deficient mice.19”
Quote in the paperNot a substantial claim0 of 1 sources checked- cites [19] Rayner KJ Sheedy FJ Esau CC Hussain FN Temel RE Parathath S van Gils JM Rayner AJ Chang AN Suarez Y Fernandez-Hernando C Fisher EA Moore KJ Antagonism of miR-33 in mice promotes reverse cholesterol transport and regression of atherosclerosis. J Clin Invest. 2011; 121: 2921- 2931 21646721 PMC3223840 source
Classified as probably not a substantial claim. May combine two claims.
PremisecitationLiver and macrophage ABCA1 both play roles in preventing atherosclerosis.
“Tissue-specific knockouts of the Abca1 locus were generated and revealed that liver and macrophage ABCA1 both play roles in preventing atherosclerosis.14”
Quote in the paperSubstantial claim0 of 1 sources checked- cites [14] Brunham LR Singaraja RR Duong M Timmins JM Fievet C Bissada N Kang MH Samra A Fruchart JC McManus B Staels B Parks JS Hayden MR Tissue-specific roles of ABCA1 influence susceptibility to atherosclerosis. Arterioscler Thromb Vasc Biol. 2009; 29: 548- 554 19201688 source
Probably not a substantial claim: may not pass S3 (load-bearing).
Study design and conductnone givenAll experimental protocols were approved by the Ethics Committee for Animal Experiments of Kyoto University.
“All the experimental protocols were approved by the Ethics Committee for Animal Experiments of Kyoto University.”
Quote in the paperNot a substantial claimClassified as probably not a substantial claim.
Study design and conductnone givenmiR-33+/+Apoe−/− littermates were used as controls.
“MiR-33+/+Apoe−/− littermates were used as controls.”
Quote in the paperSubstantial claimMay combine two claims.
Interpretationown datamiR-33 deficiency elevates serum cholesterol efflux capacity, possibly through elevation of HDL-C levels.
“These results indicated that deficiency of miR-33 elevated serum cholesterol efflux capacity, possibly through the elevation of HDL-C levels.”
Quote in the paperSubstantial claimClassified as probably not a substantial claim. May combine two claims.
Study design and conductown dataHematologic chimerism after bone marrow transplantation was determined by PCR and by measuring miR-33 levels in bone marrow cells.
“The hematologic chimerism of mice was determined by PCR using genomic DNA from blood, BM, and tail with the PCR primers described above and by measuring miR-33 levels in BM cells by quantitative PCR at age 24 weeks (ie, 16 weeks after BMT).”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
InterpretationreasoningThe real targets of miR-33 in vivo can only be clarified by genetic deletion of miR-33.
“the real targets of miR-33 in vivo can only be clarified by the genetic deletion of miR-33”
Quote in the paperSubstantial claimClassified as probably not a substantial claim.
Study design and conductreasoningAll mice used for BMT had an Apoe−/− background.
“Thus, all the mice used for BMT had an Apoe−/− background.”
Quote in the paperSubstantial claimProbably not a substantial claim: may not pass S4 (based).
InterpretationreasoningThe rise in HDL-C levels is important in the prevention of atherosclerosis.
“this may simply indicate that the rise in HDL-C levels is important in the prevention of atherosclerosis.”
Quote in the paperNot a substantial claimProbably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.
Interpretationnone givenThere seems to be a substantial difference in the effect of antisense oligonucleotide against miRNA depending on the modification, and many additional miRNAs that share an identical or similar sequence may be inactivated.
“There also seems to be a substantial difference in the effect of antisense oligonucleotide against miRNA depending on the modification such as LNA based or cholesterol modified, and many additional miRNAs that share an identical or similar sequence may be inactivated.”
Quote in the paperNot a substantial claim
The paper relies on this without support.
The ApoE−/− mouse model of atherosclerosis is a valid model for human atherosclerosis.
“In the present study, we crossed miR-33-deficient mice (miR-33−/−) with apoE-deficient mice (Apoe−/−) to examine the impact of miR-33 deletion on the progression of atherosclerosis”
The paper relies on this without support.
Cholesterol efflux measured ex vivo from macrophages reflects macrophage cholesterol efflux in atherosclerotic plaque in vivo.
“To test the hypothesis that enhanced cholesterol efflux in macrophages contributed to the reduction in the development of atherosclerosis in miR-33−/−Apoe−/− mice, cholesterol efflux to apoA-I and HDL-C was measured using 3H-cholesterol-labeled acetylated low-density lipoprotein (acLDL).”
The paper relies on this without support.
Bone marrow transplantation specifically isolates the effect of leukocyte miR-33 on atherosclerotic plaque lipid content.
“To elucidate the roles of miR-33 in blood cells, bone marrow transplantation was performed in these mice.”
The paper relies on this without support.
Apoptosis of macrophages in advanced plaques may lead to plaque rupture.
“It is generally considered that apoptosis of macrophages in advanced plaques may lead to plaque rupture.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
Inhibition or genetic ablation of miR-33a promotes regression of atherosclerosis or prevents its formation in mouse models such as LDL receptor-deficient or ApoE-deficient mice, conferring resistance to the disease.
“Inhibition or genetic ablation of miR-33a has been shown to either promote the regression of atherosclerosis or prevent its formation in mouse models, such as LDL receptor-deficient or apoprotein E (ApoE)-deficient mice, conferring resistance to the disease 21 - 23) .”
From Critical Role of microRNA-33a/b in Cardiovascular and Metabolic Disease: Molecular Mechanisms and Therapeutic Perspectives (2026) · cites this paper as [22]
The Introduction reports that antisense inhibition of miR-33 promoted plaque regression in LDL-receptor-deficient mice, and the Discussion reports smaller plaques in miR-33-deficient ApoE-deficient mice. But the paper describes the latter result as reduced progression of advanced atherosclerosis, not prevention of plaque formation or complete resistance to the disease. No page numbers are available in our copy. (LLM, conf=0.86)
Experimental — accuracy not yet measuredMacrophages within atherosclerotic lesions of these mice exhibit an anti-inflammatory phenotype characterized by enhanced cholesterol efflux, leading to a reduction in foam cell formation.
“Furthermore, macrophages within the atherosclerotic lesions of these mice exhibit an anti-inflammatory phenotype characterized by enhanced cholesterol efflux, which leads to a reduction in foam cell formation.”
From Critical Role of microRNA-33a/b in Cardiovascular and Metabolic Disease: Molecular Mechanisms and Therapeutic Perspectives (2026) · cites this paper as [22]
The Results section on peritoneal macrophages (page unavailable) reports enhanced cholesterol efflux in miR-33-deficient ApoE−/− mice, and the Discussion reports reduced plaque lipid content. But efflux was measured in peritoneal, not lesional, macrophages; the paper does not establish reduced foam-cell formation as its consequence. The Results and Discussion describe mixed pro- and anti-inflammatory markers rather than a straightforward anti-inflammatory macrophage phenotype. (LLM, conf=0.88)
Experimental — accuracy not yet measuredBone marrow transplant from miR-33 deficient animals into Apoe−/− mice reduces lipid content in atherosclerotic plaques but does not have a significant impact on plaque size.
“Additionally, bone marrow transplant from miR-33 deficient animals into Apoe−/− mice, to selectively remove miR-33 from Mθs and other hematopoietic cells, was found to reduce lipid content in atherosclerotic plaques but did not have a significant impact on plaque size (Horie et al., 2012).”
From Genetic dissection of the impact of miR-33a and miR-33b during the progression of atherosclerosis (2017) · cites this paper as Horie et al., 2012
Whole body loss of miR-33 reduces atherosclerotic plaque size and lipid content in Apoe−/− mice.
“By crossing these animals into the Apoe−/− mouse model of atherosclerosis they were able to demonstrate that whole body loss of miR-33 reduces atherosclerotic plaque size and lipid content (Horie et al., 2012).”
From Genetic dissection of the impact of miR-33a and miR-33b during the progression of atherosclerosis (2017) · cites this paper as Horie et al., 2012
These effects are contrary to what was previously reported in Apoe−/− mice lacking miR-33, where they did find reduced plaque size, did not observe any difference in circulating VLDL-C, LDL-C, or TAGs, and did not provide any data on BW or insulin sensitivity.
“These effects are contrary to what was previously reported in Apoe−/−mice lacking miR-33, where they did find reduced plaque size, did not observe any difference in circulating VLDL-C, LDL-C, or TAGs, and did not provide any data on BW or insulin sensitivity (Horie et al., 2012).”
From Genetic dissection of the impact of miR-33a and miR-33b during the progression of atherosclerosis (2017) · cites this paper as Horie et al., 2012
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Annexin V fluorophore differs between methods and figure
MinorThe methods specify Alexa Fluor 488-conjugated annexin V, but the Figure 11 legend identifies the staining reagent as Alexa Fluor 448-conjugated annexin V.
“Representative microscopic images of Alexa Fluor 448–conjugated annexin V staining.”
“staining with Alexa Fluor 488–conjugated annexin V (green)”
iNOS result has conflicting significance thresholds
MinorThe results report P=0.0057 for the iNOS-positive area, while the Figure 7 legend marks the same result as P<0.001. A P value of 0.0057 does not meet that threshold.
“the iNOS-positive area in atherosclerotic plaque in miR-33−/−Apoe−/− mice was significantly less than that in miR-33+/+Apoe−/− mice (P=0.0057).”
“B, Quantification of the iNOS-positive area in cross-sections of proximal aorta in male mice. Values are mean±SE (n=10 each); ***P<0.001.”
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Ouimet, M., Ediriweera, H. N., Gundra, U. M., Sheedy, F. J., Ramkhelawon, B., Hutchison, S. B., Rinehold, K., van Solingen, C., Fullerton, M. D., Cecchini, K., et al. (2015) · Journal of Clinical Investigation · doi:10.1172/jci81676 · Cited by 394 · see citing works
HDL-targeted therapies: progress, failures and future
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MicroRNA-33 regulates sterol regulatory element-binding protein 1 expression in mice
Horie, T., Nishino, T., Baba, O., Kuwabara, Y., Nakao, T., Nishiga, M., Usami, S., Izuhara, M., Sowa, N., Yahagi, N., et al. (2013) · Nature Communications · doi:10.1038/ncomms3883 · Cited by 230 · see citing works
Self-citationKarunakaran, D., Thrush, A. B., Nguyen, M.-A., Richards, L., Geoffrion, M., Singaravelu, R., Ramphos, E., Shangari, P., Ouimet, M., Pezacki, J. P., et al. (2015) · Circulation Research · doi:10.1161/circresaha.117.305624 · Cited by 188 · see citing works
Goedeke, L., Salerno, A., Ramírez, C. M., Guo, L., Allen, R. M., Yin, X., Langley, S. R., Esau, C., Wanschel, A., Fisher, E. A., et al. (2014) · EMBO Molecular Medicine · doi:10.15252/emmm.201404046 · Cited by 145 · see citing works
Genetic dissection of the impact of miR-33a and miR-33b during the progression of atherosclerosis
Price, N. L., Rotllan, N., Canfrán-Duque, A., Zhang, X., Pati, P., Arias, N., Moen, J., Mayr, M., Ford, D. A., Baldán, Á., et al. (2017) · Cell reports · doi:10.1016/j.celrep.2017.10.023 · Cited by 129 · see citing works
Marquart, T. J., Wu, J., Lusis, A. J., & Baldán, Á. (2013) · Arteriosclerosis, Thrombosis, and Vascular Biology · doi:10.1161/atvbaha.112.300639 · Cited by 110 · see citing works
Horie, T., Nishino, T., Baba, O., Kuwabara, Y., Nakao, T., Nishiga, M., Usami, S., Izuhara, M., Nakazeki, F., Ide, Y., et al. (2014) · Scientific Reports · doi:10.1038/srep05312 · Cited by 49 · see citing works
Self-citationMicroRNAs and Lipoprotein Metabolism
Horie, T., Baba, O., Kuwabara, Y., Yokode, M., Kita, T., Kimura, T., & Ono, K. (2014) · Journal of Atherosclerosis and Thrombosis · doi:10.5551/jat.20859 · Cited by 29 · see citing works
Self-citationBaba, O., Horie, T., Nakao, T., Hakuno, D., Nakashima, Y., Nishi, H., Kuwabara, Y., Nishiga, M., Nishino, T., Ide, Y., et al. (2018) · Molecular and Cellular Biology · doi:10.1128/mcb.00604-17 · Cited by 12 · see citing works
Self-citationHorie, T., Baba, O., Nishino, T., Yamashita, Y., Tsujisaka, Y., & Ono, K. (2026) · Journal of Atherosclerosis and Thrombosis · doi:10.5551/jat.rv22045 · Cited by 5 · see citing works
Self-citation
- Authors
- Horie, T., Baba, O., Kuwabara, Y., Chujo, Y., Watanabe, S., Kinoshita, M., Horiguchi, M., Nakamura, T., Chonabayashi, K., Hishizawa, M., Hasegawa, K., Kume, N., Yokode, M., Kita, T., Kimura, T., & Ono, K.
- Affiliations
- Kyoto University (JP)
- Kansai Medical University (JP)
- Kyoto Medical Center (JP)
- Kobe City Medical Center General Hospital (JP)
- ORCID
- Venue
- Journal of the American Heart Association
- Published
- 2012-11-29
- Volume
- 1
- Issue
- 6
- Pages
- e003376-e003376
- ISSN
- 2047-9980 (ISSN-L)
- PMC
- PMC3540673
- OpenAlex
- W2147537648
- Semantic Scholar
- 527ecb954f92f152a4810e79af124757bdbde0f6
- Cited by
- 220
- References
- 50
- Citations by year
- 2026: 3 · 2025: 8 · 2024: 7 · 2023: 10 · 2022: 10 · 2021: 14 · 2020: 17 · 2019: 15 · 2018: 19 · 2017: 14 · 2016: 26 · 2015: 26 · 2014: 28 · 2013: 23
- FWCI
- 23.19
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Locations
- Journal of the American Heart Association (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (submittedVersion, cc-by)
- Funders
- Japan Society for the Promotion of Science (23390211)
- Topics
- Health Sciences › Medicine › Surgery › Cholesterol and Lipid Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › MicroRNA in disease regulation
- Life Sciences › Immunology and Microbiology › Immunology › Atherosclerosis and Cardiovascular Diseases
- Keywords
- miR-33, atherosclerosis, atherosclerotic plaque, ABCA1, ABCG1, cholesterol efflux, HDL cholesterol, apolipoprotein A-I, macrophages, ApoE-deficient mice, RIP140, bone marrow transplantation
- MeSH terms
- ATP Binding Cassette Transporter, Subfamily G, Member 1, Animals, Apolipoproteins E, Cells, Cultured, Cholesterol, Lipoproteins, Cholesterol, HDL, Blotting, Western, Bone Marrow Transplantation, Macrophages, Peritoneal, DNA Primers, Mice, Knockout, Disease Progression, ATP-Binding Cassette Transporters, MicroRNAs, Atherosclerosis, Mice, Plaque, Atherosclerotic, Real-Time Polymerase Chain Reaction, ATP Binding Cassette Transporter 1
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-20.
Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-20.
Cited sources are checked by gpt-6-sol.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
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