Investigating the impact of CD147 and its expression on neurodegenaration and Alzheimer's disease

311 Pages·2017·4.75 MB·English

EEddiitthh CCoowwaann UUnniivveerrssiittyy
RReesseeaarrcchh OOnnlliinnee
Theses: Doctorates and Masters Theses
2012
IInnvveessttiiggaattiinngg tthhee iimmppaacctt ooff CCDD114477 aanndd iittss eexxpprreessssiioonn oonn
nneeuurrooddeeggeennaarraattiioonn aanndd AAllzzhheeiimmeerr''ss ddiisseeaassee ((AADD))
Limbikani J. Kanyenda
Edith Cowan University
Follow this and additional works at: https://ro.ecu.edu.au/theses
Part of the Medical Pathology Commons, Nervous System Diseases Commons, and the Neurology
Commons
RReeccoommmmeennddeedd CCiittaattiioonn
Kanyenda, L. J. (2012). Investigating the impact of CD147 and its expression on neurodegenaration and
Alzheimer's disease (AD). https://ro.ecu.edu.au/theses/499
This Thesis is posted at Research Online.
https://ro.ecu.edu.au/theses/499
Edith Cowan University 

Copyright Warning 

You may print or download ONE copy of this document for the purpose 
of your own research or study. 

The University does not authorize you to copy, communicate or 
otherwise make available electronically to any other person any 
copyright material contained on this site. 

You are reminded of the following: 

 Copyright owners are entitled to take legal action against persons 
who infringe their copyright. 

 A reproduction of material that is protected by copyright may be a 
copyright infringement. Where the reproduction of such material is 
done without attribution of authorship, with false attribution of 
authorship or the authorship is treated in a derogatory manner, 
this may be a breach of the author’s moral rights contained in Part 
IX of the Copyright Act 1968 (Cth). 

 Courts have the power to impose a wide range of civil and criminal 
sanctions for infringement of copyright, infringement of moral 
rights and other offences under the Copyright Act 1968 (Cth). 
Higher penalties may apply, and higher damages may be awarded, 
for offences and infringements involving the conversion of material 
into digital or electronic form
.
Investigating the impact of CD147 and its expression on
neurodegeneration and Alzheimer’s disease AD

Limbikani Jetex Thangato Kanyenda
Dip.MLT; B.Sc.; M.Sc. Human Biology

This thesis is presented in fulfilment of the requirements for
the degree of Doctor of Philosophy - Human Biology

Faculty of Computing Health and Science; School of Medical
Sciences
Edith Cowan University

Student Number: 2025800
May 2012
USE OF THESIS

The Use of Thesis statement is not included in this version of the thesis.
ABSTRACT
CD147, also known as basigin, extracellular matrix metalloproteinase inducer,
neurothelin, tumour cell-derived collagenase stimulatory factor, M6, HT7, OX47 or
gp42, is a transmembrane glycoprotein of the immunoglobulin super-family. It is
expressed in many neuronal and non-neuronal tissues with high expression in the
hippocampus, pre-frontal cortex, thyroid, heart, early erythroid, amygdala and placenta.
This protein is involved in various cellular and biological functions such as lymphocyte
migration and maturation, tissue repair, cancer progression, T and B lymphocyte
activation and induction of extracellular matrix metalloproteinase. The CD147 protein
interacts with with cyclophilin A, cyclophilin B, sterol carrier protein, caveolin-1 and
integrins, and can influence beta amyloid peptide levels, a protein that is central to
Alzheimer’s disease pathology.

Mechanisms through which CD147 regulates bata amyloid levels still remain unclear,
thus the current study investigated the impact of high cholesterol and beta amyloid 42
loading on CD147 expression in rat primary cortical neuronal cultures. Its expression in
human brain tissue from Alzheimer’s disease and non-Alzheimer’s disease dementias
and APPswe transgenic mice fed on a high fat/high cholesterol diet were also
determined. CD147 over expression experiments in rat primary cortical neuronal and
SH-SY5Y cultures using recombinant adenoviruses (AdRSV-CD147 and AdRSV-
Empty) in the presence and/or absence of cyclophilin A were employed to determine
level of proection against oxidative stress and toxicity. Finally sequencing of the CD147
promoter region (-392 to -242 nucleotides) was perfomed to identify possible single
neucrotide polymorphysims that may be linked to Alzheimer’s disease pathology.

iii
The in vitro experiments conducted in this study have shown that cholesterol and beta
amyloid loading and oxidative stress result in increased CD147 expression in rat
primary cortical neuronal cultures and that CD147 protects neuronal cultures from beta
amyloid toxicity only in the presence of cylophilin A. However in vivo experiments in
animal model have shown that a high fat/high cholesterol diet does not affect brain
CD147 levels as evidenced by unchanged expression levels in APPswe mice.

Human studies have revealed a correlation between CD147 and γ-secretase complex
components (nicastrin and presenilin 1) in the hippocampus. However, there is no
difference in CD147 protein expression between hippocampus and frontal cortex from
Alzheimer’s disease patients. Sequencing of the CD147 promoter region, where sterol
carrier protein binds to initiate CD147 transcription, has identified two novel single
neucrotide polymorphysims (-61A>G and 37C>G). However, single neucrotide
polymorphysims within this region are not associated with Alzheimer’s disease
pathology.

iv
ABBREVIATIONS
Aβ Amyloid beta peptide
AD Alzheimer’s disease
ADAM A disintegrin and a metalloproteinase
ANOVA Analysis of variance
APH1 Anterior pharynx defective homolog-1
APOE Apolipoprotein E (gene)
ApoE Apolipoprotein E (protein)
APS Ammonium persulphate
APP Amyloid precursor protein
APP-CTF Amyloid precursor protein C-terminal fragment
APH Anterior pharynx defective
ARC Animal resources centre
3β-HSD 3β-hydroxysteroid dehydrogenase
β-CTF βeta C-terminal fragment
BACE Beta amyloid cleaving enzyme
BBB Blood-brain barrier
CA1 Corn ammonis 1
CAA Congophillic amyloid angiopathy
CyPA Cyclophilin A
CyPB Cyclophilin B
o C Degrees Celsius
CSF Cerebrospinal fluid
CNS Central nervous system
CT Computed tomography
v
CTF C-terminal fragment
DDW Double de-ionised water
df Degrees of freedom
DHEA Dehydroepiandrosterone
DMEM Dulbecco’s modified Eagles medium
DFCS Dialysed foetal calf serum
DMSO Dimethyl sulphoxide
DNA Deoyxribose Nucleic Acid
DTT Dithiotreitol
ECL Enhanced chemiluminescence
EDTA Ethylenediamine tetra-acetic acid
ELISA Enzyme-linked immunosorbent assay
EMSA Eletrophoretic mobility shift assay
EMMPRIN Extracellular Matrix Metalloproteinase-Inducer
EOAD Early-onset Alzheimer’s disease
ERK1/2
FAD EFaxmtrialciaell lAullzahr-eriemgeurl’as tdeids ekaisnea se 1/2
FCS Foetal calf serum
FL-APP Full length amyloid precursor protein
HBSS Hanks balanced salt solution
HDL High density lipoprotein
HRP Horse radish peroxide
Ig Immunoglobulin
IgSF Immunoglobulin super-family
IMAC immobilized metal ion affinity chromatography
KDa Kilo Dalton
KPI Kunitz protease inhibitor
vi
LDH Lactate dehydrogenase
LDL Low density lipoprotein
LOAD Late-onset Alzheimer’s disease
LRP Low density lipoprotein receptor related protein
MAPK mitogen-activated protein kinase
MCI Mild cognitive impairment
MCT Monocarboxylate transporter
MLR Multiple linear regression
MMP Matrix metalloproteinase
MMSE Mini-Mental State Examination
MRI Magnetic resonance imaging
mRNA Messenger ribonucleic acid
MRS Magnetic resonance spectroscopy
MW Molecular weight
NATs Natural anti-sense transcripts
NCT Nicastrin
NFT Neurofibrillary tangles
NINCDS National Institute of Neurological Disorders and Communitive Disorders
and Stroke
NGF Nerve growth factor
NP40 Ethylphenyl-polyethylene glycol
PBS Phosphate buffered saline
PMSF Pheynlmethyle-sulfonyl fluoride
PBST Phosphate buffered saline with 0.05% Tween 20
PCR Polymerase chain reaction
PEN-2 Presenilin enhancer 2
PET Positron emission tomography
vii
Pg Pico gram
PHF Paired helical filaments
PS1 Presenilin 1
PS2 Presenilin 2
RIP Regulate intramembrane proteolysis
RNA Ribonucleic acid
RNAi Ribonucleic acid inhibition
ROS Reactive oxygen species
RPM Revolutions per minute
RT Room temperature
SCP Sterol carrier protein
SD Standard deviation
SDS Sodium dodecylsulfate
SE Standard error
SEM Standard error of mean
SLE Systemic lupus erythematous
SPE Sperm integral membrane proteins
StAR Steroidogenic acute regulatory protein
TACE Tumour factor-α converting enzyme
TBS Tris buffered saline
TBST Tris buffered saline with 0.05% Tween 20
TCSF Tumour cell-derived collagenase stimulatory factor
TEMED N-Tetra-methylethylenediamine
TG Triglycerides
TGFβ Transforming growth factor beta
TM Transmembrane
TMB Tetramethyl-benzidine
viii