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Cited by 55Alzheimer's disease research and treatmentsCellular transport and secretionDementia and Cognitive Impairment Research

Decreasing the expression of PICALM reduces endocytosis and the activity of β-secretase: implications for Alzheimer’s disease

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Thomas, Rhian S., Henson, Alex, Gerrish, Amy, Jones, Lesley, Williams, Julie, Kidd, Emma J.·2016·BMC Neuroscience
DOI 10.1186/s12868-016-0288-1PMC PMC4949774
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Abstract

BACKGROUND: Polymorphisms in the gene for phosphatidylinositol binding clathrin assembly protein (PICALM), an endocytic-related protein, are associated with a small, increased risk of developing Alzheimer's disease (AD), strongly suggesting that changes in endocytosis are involved in the aetiology of the disease. We have investigated the involvement of PICALM in the processing of amyloid precursor protein (APP) to understand how PICALM could be linked to the development of AD. We used siRNA to deplete levels of PICALM, its isoforms and clathrin heavy chain in the human brain-derived H4 neuroglioma cell line that expresses endogenous levels of APP. We then used Western blotting, ELISA and immunohistochemistry to detect intra- and extracellular protein levels of endocytic-related proteins, APP and APP metabolites including β-amyloid (Aβ). Levels of functional endocytosis were quantified using ALEXA 488-conjugated transferrin and flow cytometry as a marker of clathrin-mediated endocytosis (CME). RESULTS: Following depletion of all the isoforms of PICALM by siRNA in H4 cells, levels of intracellular APP, intracellular β-C-terminal fragment (β-CTF) and secreted sAPPβ (APP fragments produced by β-secretase cleavage) were significantly reduced but Aβ40 was not affected. Functional endocytosis was significantly reduced after both PICALM and clathrin depletion, highlighting the importance of PICALM in this process. However, depletion of clathrin did not affect APP but did reduce β-CTF levels. PICALM depletion altered the intracellular distribution of clathrin while clathrin reduction affected the subcellular pattern of PICALM labelling. Both PICALM and clathrin depletion reduced the expression of BACE1 mRNA and PICALM siRNA reduced protein levels. Individual depletion of PICALM isoforms 1 and 2 did not affect APP levels while clathrin depletion had a differential effect on the isoforms, increasing isoform 1 while decreasing isoform 2 expression. CONCLUSIONS: The depletion of PICALM in brain-derived cells has significant effects on the processing of APP, probably by reducing CME. In particular, it affects the production of β-CTF which is increasingly considered to be an important mediator in AD independent of Aβ. Thus a decrease in PICALM expression in the brain could be beneficial to slow or prevent the development of AD.

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  • The risk alleles of rs3851179 and rs10792832 lead to lower expression of PICALM isoform 2 in the frontal and temporal cortex in the human brain on eQTLs.

    “the risk alleles of rs3851179 (236) and rs10792832 (344) lead to lower expression of PICALM isoform 2 in the frontal and temporal cortex in the human brain on the expression quantitative trait loci (eQTLs) (586), indicating that lower PICALM levels may increase the risk for AD.”

    From Blood-Brain Barrier: From Physiology to Disease and Back (2019) · cites this paper as [586]

    The Results section, under “Association of PICALM GWAS SNPs with PICALM isoform expression,” reports that both risk alleles are associated with significantly lower isoform 2 mRNA expression in the frontal and temporal cortex in human-brain eQTL datasets. It reports an association, not evidence that the alleles cause the lower expression implied by “lead to.” (LLM, conf=0.82)

    Experimental — accuracy not yet measured

  • BIN1, CD2AP and PICALM modulate APP metabolism and the subsequent increase in Aβ production.

    “Based on pathway enrichment analyses, three GWAS-defined genes linked with endocytosis (BIN1, CD2AP and PICALM) have been proposed to modulate APP metabolism and the subsequent increase in Aβ production:”

    From The new genetic landscape of Alzheimer’s disease: from amyloid cascade to genetically driven synaptic failure hypothesis? (2019) · cites this paper as [134]

    The Conclusions (page not provided) say PICALM depletion affects APP processing, supporting that part of the claim. The Results (page not provided) report that Aβ40 was unaffected, not increased. The Background identifies BIN1 as an endocytosis-related GWAS gene, but our copy does not establish that BIN1 or CD2AP modulates APP metabolism or increases Aβ production. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • PICALM underexpression decreases levels of APP catabolites and reduces APP internalisation and Aβ generation.

    “PICALM underexpression decreases the levels of several APP catabolites, such as intracellular β-C-terminal fragment (β-CTF) and secreted sAPPβ, which has been attributed to its role in clathrin-mediated endocytosis [134].”

    From The new genetic landscape of Alzheimer’s disease: from amyloid cascade to genetically driven synaptic failure hypothesis? (2019) · cites this paper as [134]

    The Results passage reports that PICALM depletion reduced intracellular β-CTF, secreted sAPPβ, and functional endocytosis, supporting the printed citing sentence. The supplied paraphrase additionally says Aβ generation fell, but the same Results passage says Aβ40 was not affected. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

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Research works printed but not linked: 50

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Authors
Thomas, R. S., Henson, A., Gerrish, A., Jones, L., Williams, J., & Kidd, E. J.
Affiliations
  • Cardiff University (GB)
  • Birmingham Women's Hospital (GB)
Corresponding author
Kidd, Emma J.
ORCID
Venue
BMC Neuroscience
Publisher
Springer Science and Business Media LLC
Published
2016-07-18
Volume
17
Issue
1
Pages
50-50
ISSN
1471-2202 (ISSN-L)
DOI
10.1186/s12868-016-0288-1
PMC
PMC4949774
OpenAlex
W2497959445
Cited by
55
References
61
Citations by year
2026: 1 · 2025: 6 · 2024: 9 · 2023: 7 · 2022: 9 · 2021: 3 · 2020: 3 · 2019: 5 · 2018: 6 · 2017: 6
FWCI
4.56
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • BMC Neuroscience (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • ORCA Online Research @Cardiff (Cardiff University) (acceptedVersion)
  • UWE Research Repository (UWE Bristol) (submittedVersion, cc-by)
  • Cronfa (Swansea University) (submittedVersion, cc-by)
Funders
Medical Research Council (MR/L010305/1)
Topics
  • Health Sciences › Medicine › Physiology › Alzheimer's disease research and treatments
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Health Sciences › Medicine › Psychiatry and Mental health › Dementia and Cognitive Impairment Research
Keywords
PICALM, Alzheimer's disease, clathrin-mediated endocytosis, APP processing, beta-secretase, BACE1, amyloid-beta, clathrin heavy chain, siRNA knockdown, genetic polymorphism
MeSH terms
Alzheimer Disease, Brain, Endocytosis, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Humans, Immunohistochemistry, Peptide Fragments, RNA, Messenger, Transferrin, Blotting, Western, Amyloid beta-Peptides, Amyloid beta-Protein Precursor, Protein Isoforms, Clathrin Heavy Chains, Monomeric Clathrin Assembly Proteins, RNA, Small Interfering, Cell Line, Tumor, Amyloid Precursor Protein Secretases

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