The Impact of Age on Liver The Impact of Age on Liver Allograft Gene Expression Allograft Gene Expression Michael B. Ishitani, MD Michael B. Ishitani, MD William J. von Liebig Transplant Center William J. von Liebig Transplant Center Mayo Eugenio Litta Children’s Hospital Mayo Eugenio Litta Children’s Hospital Mayo Medical School, Foundation and Clinic Mayo Medical School, Foundation and Clinic Rochester, Minnesota Rochester, Minnesota
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The Impact of Age on Liver Allograft Gene Expression
The Impact of Age on Liver Allograft Gene Expression. Michael B. Ishitani, MD William J. von Liebig Transplant Center Mayo Eugenio Litta Children’s Hospital Mayo Medical School, Foundation and Clinic Rochester, Minnesota. Aging: What really happens. The Aging Process. - PowerPoint PPT Presentation
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The Impact of Age on Liver The Impact of Age on Liver Allograft Gene ExpressionAllograft Gene Expression
Michael B. Ishitani, MDMichael B. Ishitani, MDWilliam J. von Liebig Transplant CenterWilliam J. von Liebig Transplant Center
Mayo Eugenio Litta Children’s HospitalMayo Eugenio Litta Children’s Hospital
Mayo Medical School, Foundation and ClinicMayo Medical School, Foundation and Clinic
Rochester, MinnesotaRochester, Minnesota
Aging: What really happens...Aging: What really happens...
The Aging ProcessThe Aging Process
• What occurs as we Age?• Anatomic/histologic• Physiologic changes• Alterations in cellular
organelles/mitochondria/DNA• Alterations in gene expression• Individual variation
• What factors can be slowed or perhaps reversed?
• “Fountain of Youth”
Aging and the LiverAging and the Liver
• What is known of Aging and the Liver?• Hans Popper: Aging and the Liver; In Progress in
Liver Disease, 1986
• Hepatocyte life span is long (years)• Liver mitochondria life span short (days)• Large functional reserve of hepatocyte mass (15-30%)• High regenerative capability
• Gross changes: • slight decrease in weight• accumulation of brown pigment (lipofuscin)• marker of age…significance unclear• decreased blood flow
Aging and the LiverAging and the Liver
• Microscopic changes:
• polyploidy• variation in mitochondrial size• decrease in ER size• increased fibrosis• accumulation of lipofuscin
• Physiology:• Alterations in enzyme activity (variable)• Normal protein synthesis (variable)• Reduction in respiratory cycle function
[cytochrome C oxidase (COX)]• Slower/lower regeneration after injury
Aging and the LiverAging and the Liver
• Drug metabolism• Decreased due to mass/blood flow• Altered enzyme function (cytochrome P450)
• Others:• Increase in mitochondrial DNA mutations• Decrease in mitochondrial DNA repair
systems• Altered telomerase function leading to cell
senescence
Age-related Changes vary by Age-related Changes vary by Organ/TissueOrgan/Tissue
• Barrazoni, Short, and Nair: Jnl Bio Chem, 2000
Rat model of aging:• Mitochondrial DNA copy number• COX transcription level• COX enzyme activity
• Compare:• Skeletal muscle• Heart• Liver
Age-related Changes vary by Age-related Changes vary by Organ/TissueOrgan/Tissue
• Mitochondrial DNA• Decreased in liver and skeletal muscle• Heart unchanged
• Does not appear livers from an older donor “wear out”• Follow-up short-term
What happens to Older donor livers over time?
• Innocenti, et. al. Aging-related changes reverse in Pediatric Liver transplant recipients of Old donor livers, AST 2001
• Light microscopy• Accumulation of lipofuscin as
intracytoplasmic inclusions/granules in perivenular hepatocytes
• Begins in second decade and progresses• Clearance by activated macrophages in the
presence of necroinflammatory activity
Measurement of Lipofuscin in Different Measurement of Lipofuscin in Different Age Groups after Liver Transplant Age Groups after Liver Transplant
• Group 1: Young liver to Young recipient (Y-Y)Group 1: Young liver to Young recipient (Y-Y)• Donor < 18 yoDonor < 18 yo• Recipient < 18 yoRecipient < 18 yo
• Group 2: Old liver to Young recipient (O-Y)Group 2: Old liver to Young recipient (O-Y)• Donor > 50 yoDonor > 50 yo• Recipient < 18 yoRecipient < 18 yo
• Group 3: Old liver to Old recipient (O-O)Group 3: Old liver to Old recipient (O-O)• Donor > 50Donor > 50• Recipient > 65 yoRecipient > 65 yo
Materials and MethodsMaterials and Methods
• Exclusion criteria:Exclusion criteria:• Survival < 6 monthsSurvival < 6 months• No follow-up biopsies at 1 yearNo follow-up biopsies at 1 year• ACR, CMVACR, CMV
• Routine protocol biopsies: Routine protocol biopsies: • Transplant, 7d, 4 m, then yearlyTransplant, 7d, 4 m, then yearly• Standard H&EStandard H&E• Histologic review: blindedHistologic review: blinded
• Grading of lipofuscin:Grading of lipofuscin:• 00 nonenone• 11 few immediate perivenular hepatocytesfew immediate perivenular hepatocytes• 2 2 all immediate perivenular hepatocytesall immediate perivenular hepatocytes• 33 50-75% of zone 3 hepatocytes50-75% of zone 3 hepatocytes• 44 > 75% of zone 3 hepatocytes> 75% of zone 3 hepatocytes
• High density oligonucleotide microarrayHigh density oligonucleotide microarray• Hu6800 GeneChip, Affymetrix; Santa Clara, CAHu6800 GeneChip, Affymetrix; Santa Clara, CA
• Snap biopsies at reperfusion and 1 year (0.5g)Snap biopsies at reperfusion and 1 year (0.5g)
• Comparisons made between 1h and 1y in each Comparisons made between 1h and 1y in each groupgroup
Results of Gene Chip AnalysisResults of Gene Chip Analysis
A B G ene
Im m une function/Lipid m etabolismá 3.0 (á 2.0) H LA class-I
á 3.0 (á 2.0) H LA class-II
á 4.4 (á 2.5) Prop. C oA carboxylase
á 3.6 (á 2.4) 3-Oxoacyl C oA th iolase
A = old (donor) to young (rec ipie nt); B= old (donor) to old (re cipient).
A B Gene
DNA maintenance/ Protein metabolismá 4.6 (á 2.4) Glutathione peroxidase
á 4.2 (á 2.9) Catalase
á 6.4 (á 2.3) Elongation factor-TU
á 7.3 (á 3.1) Transcription EF-SII
A= old (donor) to young (recipient); B= old (donor) to old (recipient).