US007622271 B2

(12) United States Patent (10) Patent No.: US 7.622,271 B2 Kennedy et al. (45) Date of Patent: Nov. 24, 2009

(54) IDENTIFICATION OF AGING Itahana et al. (2003) Molecular and Cellular Biology, vol. 23, No. 1, THROUGH LARGE-SCALEANALYSIS pp. 389-401.* Kapahi et al. (2004) Current Biology, vol. 14, pp. 885-890.* (75) Inventors: Brian K. Kennedy, Redmond, WA (US); Lin et al. (2000) Science. vol. 289, pp. 2126-2128.* Vellai et al. (2003) Nature. vol. 426, pp. 620-621.* Matthew R. Kaeberlein, Kirkland, WA Chen et al. Experimental Gerontology (2003) vol. 38, pp. 1051 (US) 1063. Abraham, R.T. et al., “Immunopharmacology of Rapamycin.” Annu. 73) Assignee:9. Universityty of Washington,g Seattle, WA Rev. Immuno. 14, 483, 1996. (US) Beretta, L. et al., “Rapamycin blocks the of 4E-BP1 and inhibits cap-dependent initiation of translation.” EMBO.J. 15(3), *) Notice: Subject to anyy disclaimer, the term of this 658-664, 1996. patent is extended or adjusted under 35 Brown, E. J. et al., “A mammalian targeted by Gl-arresting U.S.C. 154(b) by 510 days. rapamycin-receptor complex.” Nature 369, 756-758, Jun. 30, 1994. Dumont, F.J. et al., “Distinct Mechanisms of Supression of Murine T (21) Appl. No.: 11/107,542 Activation by the Related Macrollides FK-506 and Rapamycin.” J. Immunol 144, 251-258, 1990. (22) Filed: Apr. 15, 2005 Feldmann, H. et al., "Complete DNA sequence of yeast II, EMBO.J. 13,5795-5809, 1994. (65) Prior Publication Data (Continued) US 2006/0068414A1 Mar. 30, 2006 Primary Examiner Lori A Clow Related U.S. Application Data (74) Attorney, Agent, or Firm Fenwick & West LLP (60) Provisional application No. 60/591461, filed on Jul. (57) ABSTRACT 26, 2004. High throughput methods for screening genetic variants that (51) Int. Cl. are phenotypically distinguishable are provided. Methods for GOIN 33/53 (2006.01) identifying “long-lived genetic variants among a set of vari ADIN 63/ (2006.01) ants are also provided. Methods for identifying pharmaceu G06F 9/00 (2006.01) tical compounds that can promote longevity in various Sub (52) U.S. Cl...... 435/7.31; 424/93.1: 702/19 jects, including mammals, and that can delay the onset of (58) Field of Classification Search ...... None various diseases associated with aging are also provided. See application file for complete search history. Various vectors and host cells containing identified genes/ products are useful for screening longevity-promoting (56) References Cited compounds that can interact with life-span-regulating genes/ OTHER PUBLICATIONS gene products. Pharmaceutical compositions that can pro Chen et al. (2003) Experimental Gerontology, vol. 38, pp. 1051 mote longevity are also provided. 1063. Fabrizio et al. (2004) FEBS Letters. vol. 557, pp. 136-142.* 10 Claims, 25 Drawing Sheets Replicative Life Span (RLS) Analysis s?" Goff" M f (D2) O (M52. cell divisions prior to senescence? 202 204 206 208 US 7.622.271 B2 Page 2

OTHER PUBLICATIONS Kunz, J. et al., “Target of Rapamycin in Yeast, TOR2, Is an Essential Jefferies, H. B. J. et al., “Rapamycin suppresses 5 TOP mRNA Phosphatidylinositol Homolog Required for G Progression.” Goffeau et al., “Life with 6000 Genes. Science 274, 546-547, 1996. Cell 73,585-596, 1993. Huang, M-E, et al., “Analysis of a 62 kb DNA Sequence of Chromo Philippsen, P. et al., “The nucleotide sequence of Saccharomyces some X Reveals 36 Open Reading Frames and a Gene Cluster with a XIV and its evolutionary implications.” Nature 387(6632), 93-98, Counterpart on Chromosome XI.” Yeast 12, 869-875, 1996. transla 1997. tion through inhibition of p70.” EMBO.J. 16(12), 3693-3704, Raught, B. et al., “The target of rapamycin (TOR)