The Regulation of INK4/ARF In Cancer and Aging

William Y. Kim and Norman E. Sharpless Cell 127, October 20, 2006. Oscar Leonardo Mosquera Dussan Electronic Engineer.

Outline

Overview The INK4 Family and ARF Role in Cancer Role in Aging Regulation of Expression Silencing of the INK4a/ARF/INK4b locus

INK4a/ARF/INK4b
Decline in replicative Self-renewing Cell during aging contributes p16 Chromosome 9p21 Products P15,p16,ARF Regulation Uncontrolled Proliferation Hallmark of cancer Loss of locus Frequent event In human cancer

Rapid, Disordered, Unexpected, And invasive (Inflamatory Response to A wound )

Melanoma, Pancreatic Adenocarcinoma, Glioblastoma, Certain leukimias, Lung cancer, Bladder carcinoma

How a tumor suppressor protein What regulates the Distinguishes malignant from phisiological INK4a/ARF/INK4b locus Growth ? Should help to elucidate Properties a would-be Activate expression Cell recognize as Of the malignant INK4a/ARF/INK4b locus

The INK4 Family and ARF

In a small 35kb of the human genome the locus encodes 3 T.S.P. P16 and ARF are not isoform. INK4 class are homologous inhibitors of CDK4/6 which promotes proliferation P15 and p16 are 85% similar at the aminoacid level, expression maintains Rb proteins in a hypophosphorylated state ARF Regulate p53 in response to abberant growth or oncogenesis stress such as c-MYC activation. ARF binto to and inactivates the MDM2 protein, and M2M in turn regulates p53 as an Ubiquitin ligase for proteosomal degradation.

Physical proximity and shared sequences of The locus allow p16,ARF,p15 to sense and Coordinately respond to common stimuli in Nascent tumor.

Role in Cancer
Loss of p16, Point mutation or Small deletion Which member(s) of the Locus represent the principal Tumor suppressor activity p15 Reported in Thousand of Human cancer (Forbes et al., 2006) Stronger Case, More important In humans. Overlapping Biochemical function Co-deletion may be More oncogenic Knock-out studies Of mice All 3 strains are More prone to Spontaneous Cancer than wild Type littermates ARF

Specific lesions are Not well described Rare glial tumors and Certain leukimia

More important In mice Redundancy Acute lymphoblastic leukemia [ALL] -Deletion of 9p21 (targeting All 3 proteins)

Role in Aging
Mammalian ageing has Been associated with Reduced regenerative capacity Expression of p16 Increases markedly With aging in many tissues Work in lab P16 deficient and Overexpressing mice Suggest that P16 expression is One cause of aging Promotes by Limiting proliferation And self-renewal P16 appears capable Of promoting aging In disparate tissues That are Developmentally distinct Effects of p16 loss Were remarkably Consistent Zindy et al., 1997 Krishnamurthy et al., 2004 Melk et al., 2004 Nielsen et al., 1999

-HSCs -NSCs -Pancreatic islet (Unipotent cells) Appear to require Cdk4/6 for proliferation In no organ studied (bone marrow, brain, Endocrine pancreas) did P16 loss completely Abrogate the effects of aging

Brains from mice lacking P16 still demonstrate reduced Neurogenesis with aging

Regulation of Expression

INK4a/ARF increases at an early stage of tumorigenesis ( reviewed in sherr, 2000) , precise stimuli relevant to cancer that induce the expression of the locus are unknown The molecular characteristics of aging that lead to increased p16 expression are simlarly unclear Benefical signals vs undesirable signals. In rodents, p16 and ARF are coregulated, in human is not well established.

Silencing of the locus

DNA replication CDC6 interacts with the Origin recognition Complex and recruits Factors that unwind the DNA helix Silencing the locus

Gonzales et al, 2006 Reported a coordination

Heterochromatinization of RD induced by RNAi

They hipothesized that CDC6 expression might Regulate both, DNA Replication and Transcription at RD

Transcriptional Repression of the locus

Suggest a novel molecular connection Between DNA replication and INK4A/ARF/INK4B transcription.