CoChat
Sign Up
Cited by 165

The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.

Genome-wide RNAi screen reveals ALK1 mediates LDL uptake and transcytosis in endothelial cells

Kraehling, Jan R., Chidlow, John H., Rajagopal, Chitra, Sugiyama, Michael G., Fowler, Joseph W., Lee, Monica Y., Zhang, Xinbo, Ramírez, Cristina M., Park, Eon Joo, Tao, Bo, Chen, Keyang, Kuruvilla, Leena, Larriveé, Bruno, Folta-Stogniew, Ewa, Ola, Roxana, Rotllan, Noemi, Zhou, Wenping, Nagle, Michael W., Herz, Joachim, Williams, Kevin Jon, Eichmann, Anne, Lee, Warren L., Fernández-Hernando, Carlos, Sessa, William C.·2016·Nature Communications
DOI 10.1038/ncomms13516PMC PMC5121336
Open this paper in CoChat — free account

Abstract

In humans and animals lacking functional LDL receptor (LDLR), LDL from plasma still readily traverses the endothelium. To identify the pathways of LDL uptake, a genome-wide RNAi screen was performed in endothelial cells and cross-referenced with GWAS-data sets. Here we show that the activin-like kinase 1 (ALK1) mediates LDL uptake into endothelial cells. ALK1 binds LDL with lower affinity than LDLR and saturates only at hypercholesterolemic concentrations. ALK1 mediates uptake of LDL into endothelial cells via an unusual endocytic pathway that diverts the ligand from lysosomal degradation and promotes LDL transcytosis. The endothelium-specific genetic ablation of Alk1 in Ldlr- KO animals leads to less LDL uptake into the aortic endothelium, showing its physiological role in endothelial lipoprotein metabolism. In summary, identification of pathways mediating LDLR-independent uptake of LDL may provide unique opportunities to block the initiation of LDL accumulation in the vessel wall or augment hepatic LDLR-dependent clearance of LDL.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1038/ncomms13516

Cites

3 of 3 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

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?

  • Comparison of the authors' data to a previous siRNA screen for endothelial cell LDL uptake revealed limited overlap, with only 1 gene identified in both studies.

    “Comparison of our data to a previous siRNA screen for endothelial cell LDL uptake[19] revealed limited overlap, with only 1 gene identified in both studies (S9 Table).”

    From Genome-scale CRISPR screening for modifiers of cellular LDL uptake (2021) · cites this paper as [19]

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Flagged first, then by citations

Research works printed but not linked: 62

  1. Tabas, I., Williams, K. J. & Boren, J. Subendothelial lipoprotein retention as the initiating process in atherosclerosis: update and therapeutic implications. Circulation 116, 1832–1844 (2007). · DOI · Paper page
  2. Williams, K. J. & Tabas, I. Lipoprotein retention--and clues for atheroma regression. Arterioscler. Thromb. Vasc. Biol. 25, 1536–1540 (2005). · DOI
  3. Williams, K. J. & Tabas, I. The response-to-retention hypothesis of early atherogenesis. Arterioscler. Thromb. Vasc. Biol. 15, 551–561 (1995). · DOI · Paper page
  4. Vasile, E., Simionescu, M. & Simionescu, N. Visualization of the binding, endocytosis, and transcytosis of low-density lipoprotein in the arterial endothelium in situ. J. Cell Biol. 96, 1677–1689 (1983). · DOI · Paper page
  5. Brown, M. S. & Goldstein, J. L. Regulation of the activity of the low density lipoprotein receptor in human fibroblasts. Cell 6, 307–316 (1975). · DOI
  6. Rippe, B., Rosengren, B. I., Carlsson, O. & Venturoli, D. Transendothelial transport: the vesicle controversy. J. Vasc. Res. 39, 375–390 (2002). · DOI
  7. Kuzmenko, E. S., Djafarzadeh, S., Cakar, Z. P. & Fiedler, K. LDL transcytosis by protein membrane diffusion. Int. J. Biochem. Cell Biol. 36, 519–534 (2004). · DOI
  8. Wiklund, O., Carew, T. E. & Steinberg, D. Role of the low density lipoprotein receptor in penetration of low density lipoprotein into rabbit aortic wall. Arteriosclerosis 5, 135–141 (1985). · DOI
  9. Bartels, E. D., Christoffersen, C., Lindholm, M. W. & Nielsen, L. B. Altered metabolism of LDL in the arterial wall precedes atherosclerosis regression. Circ. Res. 117, 933–942 (2015). · DOI
  10. Edgell, C. J., McDonald, C. C. & Graham, J. B. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc. Natl Acad. Sci. USA 80, 3734–3737 (1983). · DOI
  11. David, L., Mallet, C., Mazerbourg, S., Feige, J. J. & Bailly, S. Identification of BMP9 and BMP10 as functional activators of the orphan activin receptor-like kinase 1 (ALK1) in endothelial cells. Blood 109, 1953–1961 (2007). · DOI
  12. Lin, M. I., Yu, J., Murata, T. & Sessa, W. C. Caveolin-1-deficient mice have increased tumor microvascular permeability, angiogenesis, and growth. Cancer Res. 67, 2849–2856 (2007). · DOI
  13. Heinecke, J. W. Free radical modification of low-density lipoprotein: mechanisms and biological consequences. Free Radic. Biol. Med. 3, 65–73 (1987). · DOI
  14. Goldstein, J. L., Brunschede, G. Y. & Brown, M. S. Inhibition of proteolytic degradation of low density lipoprotein in human fibroblasts by chloroquine, concanavalin A, and Triton WR 1339. J. Biol. Chem. 250, 7854–7862 (1975). · DOI
  15. Fisher, T. S. et al. Effects of pH and low density lipoprotein (LDL) on PCSK9-dependent LDL receptor regulation. J. Biol. Chem. 282, 20502–20512 (2007). · DOI · Paper page
  16. Armstrong, S. M. et al. A novel assay uncovers an unexpected role for SR-BI in LDL transcytosis. Cardiovasc. Res. 108, 268–277 (2015). · DOI
  17. Park, S. O. et al. Real-time imaging of de novo arteriovenous malformation in a mouse model of hereditary hemorrhagic telangiectasia. J. Clin. Invest. 119, 3487–3496 (2009).
  18. Tual-Chalot, S. et al. Endothelial depletion of acvrl1 in mice leads to arteriovenous malformations associated with reduced endoglin expression. PLoS ONE 9, e98646 (2014). · DOI
  19. Hobbs, H. H., Brown, M. S. & Goldstein, J. L. Molecular genetics of the LDL receptor gene in familial hypercholesterolemia. Hum. Mutat. 1, 445–466 (1992). · DOI · Paper page
  20. Lesnik, P., Haskell, C. A. & Charo, I. F. Decreased atherosclerosis in CX3CR1−/− mice reveals a role for fractalkine in atherogenesis. J. Clin. Invest. 111, 333–340 (2003). · DOI · Paper page
  21. Combadiere, C. et al. Decreased atherosclerotic lesion formation in CX3CR1/apolipoprotein E double knockout mice. Circulation 107, 1009–1016 (2003). · DOI · Paper page
  22. Poupel, L. et al. Pharmacological inhibition of the chemokine receptor, CX3CR1, reduces atherosclerosis in mice. Arterioscler. Thromb. Vasc. Biol. 33, 2297–2305 (2013). · DOI
  23. Apostolov, E. O. et al. Endonuclease G mediates endothelial cell death induced by carbamylated LDL. Am. J. Physiol. Heart Circ. Physiol. 300, H1997–H2004 (2011). · DOI
  24. Moran, C. S., Campbell, J. H. & Campbell, G. R. Human leukemia inhibitory factor upregulates LDL receptors on liver cells and decreases serum cholesterol in the cholesterol-fed rabbit. Arterioscler. Thromb. Vasc. Biol. 17, 1267–1273 (1997). · DOI
  25. Fernandez-Hernando, C. et al. Genetic evidence supporting a critical role of endothelial caveolin-1 during the progression of atherosclerosis. Cell Metab. 10, 48–54 (2009). · DOI
  26. Frank, P. G., Pavlides, S., Cheung, M. W., Daumer, K. & Lisanti, M. P. Role of caveolin-1 in the regulation of lipoprotein metabolism. Am. J. Physiol. Cell Physiol. 295, C242–C248 (2008). · DOI
  27. Frank, P. G. et al. Genetic ablation of caveolin-1 confers protection against atherosclerosis. Arterioscler. Thromb. Vasc. Biol. 24, 98–105 (2004). · DOI
  28. Calvo, D., Gómez-Coronado, D., Suárez, Y., Lasunción, M. A. & Vega, M. A. Human CD36 is a high affinity receptor for the native lipoproteins HDL, LDL, and VLDL. J. Lipid Res. 39, 777–788 (1998). · DOI
  29. Santibanez, J. F. et al. Caveolin-1 interacts and cooperates with the transforming growth factor-beta type I receptor ALK1 in endothelial caveolae. Cardiovasc. Res. 77, 791–799 (2008). · DOI
  30. Trigatti, B. et al. Influence of the high density lipoprotein receptor SR-BI on reproductive and cardiovascular pathophysiology. Proc. Natl Acad. Sci. USA 96, 9322–9327 (1999). · DOI
  31. Maisonpierre, P. C. et al. Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science 277, 55–60 (1997). · DOI · Paper page
  32. Valenzuela, D. M. et al. Angiopoietins 3 and 4: diverging gene counterparts in mice and humans. Proc. Natl Acad. Sci. USA 96, 1904–1909 (1999). · DOI
  33. Lee, H. J. et al. Biological characterization of angiopoietin-3 and angiopoietin-4. FASEB J. 18, 1200–1208 (2004). · DOI
  34. Xiao, L., Harrell, J. C., Perou, C. M. & Dudley, A. C. Identification of a stable molecular signature in mammary tumor endothelial cells that persists in vitro. Angiogenesis 17, 511–518 (2014). · DOI
  35. Takase, H. et al. Genome-wide identification of endothelial cell-enriched genes in the mouse embryo. Blood 120, 914–923 (2012). · DOI
  36. Carew, T. E., Pittman, R. C., Marchand, E. R. & Steinberg, D. Measurement in vivo of irreversible degradation of low density lipoprotein in the rabbit aorta. Predominance of intimal degradation. Arteriosclerosis 4, 214–224 (1984). · DOI
  37. Dehouck, B. et al. A new function for the LDL receptor: transcytosis of LDL across the blood-brain barrier. J. Cell Biol. 138, 877–889 (1997). · DOI · Paper page
  38. Yao, Y., Zebboudj, A. F., Torres, A., Shao, E. & Bostrom, K. Activin-like kinase receptor 1 (ALK1) in atherosclerotic lesions and vascular mesenchymal cells. Cardiovasc. Res. 74, 279–289 (2007). · DOI
  39. Yao, Y. et al. Inhibition of bone morphogenetic proteins protects against atherosclerosis and vascular calcification. Circ. Res. 107, 485–494 (2010). · DOI
  40. Derwall, M. et al. Inhibition of bone morphogenetic protein signaling reduces vascular calcification and atherosclerosis. Arterioscler. Thromb. Vasc. Biol. 32, 613–622 (2012). · DOI
  41. Oh, S. P. et al. Activin receptor-like kinase 1 modulates transforming growth factor-beta 1 signaling in the regulation of angiogenesis. Proc. Natl Acad. Sci. USA 97, 2626–2631 (2000). · DOI
  42. Brautbar, A. & Ballantyne, C. M. Pharmacological strategies for lowering LDL cholesterol: statins and beyond. Nat. Rev. Cardiol. 8, 253–265 (2011). · DOI
  43. Birmingham, A. et al. Statistical methods for analysis of high-throughput RNA interference screens. Nat. Methods 6, 569–575 (2009). · DOI · Paper page
  44. Goedeke, L. et al. MicroRNA-148a regulates LDL receptor and ABCA1 expression to control circulating lipoprotein levels. Nat. Med. 21, 1280–1289 (2015). · DOI · Paper page
  45. Ghosh, R. N., Gelman, D. L. & Maxfield, F. R. Quantification of low density lipoprotein and transferrin endocytic sorting HEp2 cells using confocal microscopy. J. Cell Sci. 107, 2177–2189 (1994). · DOI · Paper page
  46. Allport, J. R. et al. Neutrophils from MMP-9- or neutrophil elastase-deficient mice show no defect in transendothelial migration under flow in vitro. J. Leukoc. Biol. 71, 821–828 (2002). · DOI
  47. Park, S. O. et al. ALK5- and TGFBR2-independent role of ALK1 in the pathogenesis of hereditary hemorrhagic telangiectasia type 2. Blood 111, 633–642 (2008). · DOI
  48. Benedito, R. et al. The notch ligands Dll4 and Jagged1 have opposing effects on angiogenesis. Cell 137, 1124–1135 (2009). · DOI
  49. Ricard, N. et al. Functional analysis of the BMP9 response of ALK1 mutants from HHT2 patients: a diagnostic tool for novel ACVRL1 mutations. Blood 116, 1604–1612 (2010). · DOI
  50. Idevall-Hagren, O., Dickson, E. J., Hille, B., Toomre, D. K. & De Camilli, P. Optogenetic control of phosphoinositide metabolism. Proc. Natl Acad. Sci. USA 109, E2316–E2323 (2012). · DOI
  51. Harrison, K. D. et al. Nogo-B receptor stabilizes Niemann-Pick type C2 protein and regulates intracellular cholesterol trafficking. Cell Metab. 10, 208–218 (2009). · DOI
  52. Goldstein, J. L., Basu, S. K. & Brown, M. S. Receptor-mediated endocytosis of low-density lipoprotein in cultured cells. Methods Enzymol. 98, 241–260 (1983). · DOI
  53. Williams, K. J. in Proteoglycan Protocols Vol. 171. ed. Iozzo Renato V. Ch. 45 457–477Springer Science & Business Media (2001).
  54. Sobal, G., Resch, U. & Sinzinger, H. Modification of low-density lipoprotein by different radioiodination methods. Nucl. Med. Biol. 31, 381–388 (2004). · DOI
  55. Vieira, O. V., Laranjinha, J. A., Madeira, V. M. & Almeida, L. M. Rapid isolation of low density lipoproteins in a concentrated fraction free from water-soluble plasma antioxidants. J. Lipid Res. 37, 2715–2721 (1996). · DOI
  56. Pitas, R. E., Innerarity, T. L., Weinstein, J. N. & Mahley, R. W. Acetoacetylated lipoproteins used to distinguish fibroblasts from macrophages in vitro by fluorescence microscopy. Arteriosclerosis 1, 177–185 (1981). · DOI
  57. Teupser, D., Thiery, J., Walli, A. K. & Seidel, D. Determination of LDL- and scavenger-receptor activity in adherent and non-adherent cultured cells with a new single-step fluorometric assay. Biochimica et biophysica acta 1303, 193–198 (1996). · DOI
  58. Azizi, P. M. et al. Clathrin-dependent entry and vesicle-mediated exocytosis define insulin transcytosis across microvascular endothelial cells. Mol. Biol. Cell 26, 740–750 (2015). · DOI
  59. Roussel, F. & Dalion, J. Lectins as markers of endothelial-cells - comparative-study between human and animal-cells. Lab. Anim. 22, 135–140 (1988). · DOI
  60. Carpenter, A. E. et al. Cell Profiler: image analysis software for identifying and quantifying cell phenotypes. Genome Biol. 7, R100 (2006). · DOI
  61. Kamentsky, L. et al. Improved structure, function and compatibility for CellProfiler: modular high-throughput image analysis software. Bioinformatics 27, 1179–1180 (2011). · DOI
  62. Börnig, H. & Geyer, G. Staining of cholesterol with the fluorescent antibiotic ‘filipin’. Acta histochemica 50, 110–115 (1974).

Cited by

Sorted by citations

Metadata

Authors
Kraehling, J. R., Chidlow, J. H., Rajagopal, C., Sugiyama, M. G., Fowler, J. W., Lee, M. Y., Zhang, X., Ramírez, C. M., Park, E. J., Tao, B., Chen, K., Kuruvilla, L., Larriveé, B., Folta-Stogniew, E., Ola, R., Rotllan, N., Zhou, W., Nagle, M. W., Herz, J., Williams, K. J., Eichmann, A., Lee, W. L., Fernández-Hernando, C., & Sessa, W. C.
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2016-11-21
Volume
7
Issue
1
ISSN
2041-1723
DOI
10.1038/ncomms13516
PMC
PMC5121336
Cited by
165
References
3
Language
en
Licence

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.

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.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review