G(-670)-> A. polymophism in the promoter of the fas gene is a risk factor for myocardial infarction --Miyuki Onishi et al.

1
G(-670)-gt A. polymophism in the promoter of the
fas gene is a risk factor for myocardial
infarction --Miyuki Onishi et al.

• Fas is a receptor that mediates apoptosis which
  is a critical event for eliminating activated
  macrophages.
• Method
• 154 subject with recent MI, 462 control.
• Fas expression was detected on peripheral
  blood mononuclear cells.
• Results
• A/G polymorphism of the fas gene
  significantly
• different between MI and controls.
• Leukemia inhibitory factor
  significantly suppressed the
• levels of Fas expression in AA
  genotype subjects, no
• effect in subjects with GG genotype.
• Conclusion The A allele of G/A polymorphysm in
  the promoter region (-670-A) of the Fas gene is a
  new genetic risk factor of MI via modulation of
  Fas expression

2
Immunoglobulin Treatment Suppresses
Atherosclerosis in Apolipoprotein E Knockout Mice
via the FC portion--Zhuyi Yuan et al

• IG is used for the treatment of immune-mediated
  disease.
• Method
• ApoE mice fed with high fat diet.
• IP injection of Human IG or F(ab)2
• fragments of IG at 8 weeks and 16
  weeks
• Results
• IG markedly suppressed Fatty streak
  formation
• and fibofatty plaque and macrophage
  accumulation
• F(ab)2 has no effect.
• Conclusion
• IG therapy markedly suppressed
  atherosclerosis via the Fc
• portion of IG.
• The suppression is associated with
  marked reduction
• of macrophage density in the region.

3
NF-kB Inhibition Reduces Pro-Inflammatory
Molecule Expression and Attenuates
Atherosclerosis in ApoE-/- miceYutaka Furukawa
et al.

• Activation of NF-kB induces pro-inflammatory
  molecule mRNA including MCP-1 and VCAM-1.
• Method
• NF-KP inhibitor(NF-kB1) was used.
• Male C57BL/6 apoE-/- mice were fed a
  western type diet.
• Administ. Of NF-kB1 or vehicle for 10
  weeks.
• Results
• Immunohistochemistry macrophage
  accumulation and VCAM-1
• expression attenuated.
• Real-time PCR showed that VCAM-1 and
  MCP-1 mRNA
• expression in the aorta decreased
• Conclution
• NF-kB may be a therapeutic target in
  the prevention of
• atherosclerosis.

4
Cholesterol Regulated Genome wide approchBreslow

• Using microarray to identify genes regulated by
  dietary cholesterol in the mouse live.
• Method C57BL/6J mice

• PNAS (2002)996949-6954
• JBC(2002)2764261
• Biochemistry(2000)3915399
• Science 2901712
• Proteins(2001)43134

W/o cholestrol
With 0.5 cholesterol.
Pooled or individual hybridize with array.
Retain genes with signal gt 250, Foldgt1.6.
Unigen cluster tools, got 88 probe set
There are 15 starts in Human Genome. He
introduced Star D4,5,6 from the probe set.
5
MMP-8 Evidence of a New Collagenolytic Pathway
in Acute Plaque DisruptionLan Loftus et al.

• MMP-8 is the enzyme that preferentially degrades
  type I collagen.
• Aim quantify the level of MMP-8 in carotid
  plaques.
• Method
• Plaque from 75 patients
  undergoing carotid endarterectomy.
• MMP-8 level were quantified
  using ELISA.Immunostaining.
• All patient undergo Td
  monitoring to detect spontaneous
• embolisation.
• Results MMP-8 level in plaque higher in patients
  with recent symptoms and
• spontaneous embolisation.
• There is significant
  relation between level of MMP-8 and MMP-9.
• Conclusion
• Elevate of MMP-8 and
  together with MMP-9 have the potential to
• degrade all components of
  extracellular matrix and represent a target
• for pharmacotherapy to
  prevent acute plaque disruption.

6
Enhanced Migration and Matrix Degradation by
Chlamydia pneumoniae-Infected Monocytes via the
plasminogen and EMMPRIN/MMP Activation system.
Implications for Plaque Rupture. -- Roland
Schmidt et al.

• The effects of C.pn initiation of matrix
  degrading activity in monocytes with regard to
  the role of EMMPRIN, Extracellular MMP-inducer.
• Method and results
• MonoMac6 cells or isolated monocytes
• with C pn for 48 hrs.
• MMP-2MMP-9,uPAR,membrand-type-1-MMP,EMMPR
  IN
• expression enhanced
• Conclusion
• C.pn. Induces monocytic matrix degradation
  and matrix invasion via activation of the
  plasminogen and the EMMPRIN/MMP activative
  system. Which may contribute to the development
  of cardiovascular complications such as plaque
  rupture.

7
Angiotensin II mediates production of Matrix
Metalloproteinases-1 and 9 by Monocytes
Implication for Atherosclerotic Plaque Rupture
Min P Kim et al.

• Hypothesis AngII enhances the production of
  monocyte MMP-1 and MMP-9, two major MMPs
  associated with plaque rupture.
• Method Elutration of human primary blood
  monocytes cultured with
• various agents.
• Levels of MMP-2,MMP-9 in
  supernatant were determined by
• ELISA or western analysis.
• Conclusion AngII has a central role in the
  production of MMP-1 and MMP-9 by monocytes. It
  provide insight into the association between
  hypertension and acut coronary sydrome and a
  possible mechanism by which angiotensin
  converting enzyme inhibitor and aspirin may
  reduce the risk for heart attacks.

8
Organization of HUPO and opportunity for
proteomic research on a globe wide level as well
as in the USA Sam Hanash
Protein microarray Identify antigen in
autoimmunity. Identify drug binging
protein. Studies of protein interaction

• Why proteomics
• High dynamic compartment
• Most direct relevant to function
• High relevant to biological fluid
• Regulated at transcriptional level
• and post-transcription level.
• Field of Proteomics
• Expression protein
• Structure protein
• Protein function
• Proteomic informatics

Types of Technology Separation Basic
2D/Mass spec. present focus on subcellular
compartment. Exp Cell surface protein which is
important for diagnostics and therapeutics Tag
cell surface protein with Biotin. Then solubolize
cell membrane. Run this cell lysate through
Avidin column to capture Biotin related cell
surface protein. Load onto 2D gel for protein
separation. Digest all interest band and identify
them with Mass. Combination of
electrophoresis and chromatography to fractionate
one lysate into a lot of protein.
International cooperation 1. Reesources and
tech. 2. Model cells. 3. Plasma proteomic 4.
Informatics.
Goal Comprehensive analyze of all of protein
constituents of a cell or tissue. Tracking of
protein traffic. Tracking of post-translational
modifiers
Model cells and tissure Liver proteomic Heart
proteomic Brain proteomic Plasma proteomic.
9
Gene Expression Profiles during Human
Atherogeeneesis A Role for the LIF-Receptor in
Lesion Progression --Birgit Faber et al

• Study profle large-scale expression on early,
  advanced but stable
• and ruptured atherosclerotic
  plaques.
• Method and results
• 1. 3 early stage plaque and 3 ruptured
  plaque mRNA -gt run on microarray ( Incyte
  Genomics Inc) -gt 105 gene upregulated and 88
  genes down regulated, phase of atherogenesis vs.
  stable plaques. 79 gene up and 65 gene down after
  plaque rupture.
• 2. Hybridize CDNA of (Pooled n3 for each)
  normal artery, early lesions, stable lesions and
  rupture lesion with custome made cDNA expression
  array. LIF receptor enhanced in plaque rupture.
  RT-PCR revealed LIF receptor MRNA expression in
  1/3 normal arteries, ¼ early leesions,8/10ruptured
  plaque specimens. LIP receptor protein
  expression located in smooth muscle cells and
  macrophages of atherosclerotic lesions.
• Conclusion
• This expression profile suggests a role for
  the LIF-receptor in the progression of human
  atherosclerosis.

10
Cyr61(CNN1) and Connective Tissue Growth
Factor(CNN2) are Novel Ligands of Inteegrin aMB2
on Peeripheral Blood Monocytes -- Joseph M
Schober et al.

• Cysteine-rich 61(Cry61,CCN1) and connective
  tissuee growth factor(CTGF, CVN2) are growth
  factor-incucible immediate-early gene products
  found in atherosclerotic lesions, restenosed
  blood vellel walls and healing cutaneous wounds.
• Study Examinee monocyte interaction with Cyr61
  and CTGF. Because monocyte adhesion and
  transmigration are important for
  athereosclerosis, inflammation and wound healing.
• Results monocyte adhesion to Cyr61 is
  specificaly blocked by antibodies directed
  against the integrin aM and B2 subunit indicating
  that integrin serves as an adhesion recptor on
  monocyte for thesee CCN protein. Monocyte
  adhesion to Cyr61 induces the expression of
  inflammatory mediators including IL-1, IL-8
  MCP-1,MRP8 and MRP14.
• Suggestion these finding indicated that Cyr61
  and CTGF aree novel ligands of integrin aMB2 on
  monocytes and suggestion an important
  pathophysiologic role of monocyte adhesion to CNN
  protein.