Lombardi Cancer Center Georgetown University Washington, DC

Lombardi Cancer Center Georgetown University Washington, DC microrna and other nucleic acids analysis in liquid biopsies Anton Wellstein World CDx Boston, Sept 8-11, 2015

Disclosures: Inventor on Georgetown University filed patents related to the topic Member of the Scientific Advisory Board of Indivumed Inc.

DNA mrnas + 3 x 10 4 genes + 10 5 mrnas Proteins >10 6-10 7 modified Metabolites + 5 x 10 3 Phenotype

DNA mrnas + 3 x 10 4 genes + 10 5 mrnas + 10 3 micrornas Proteins >10 6-10 7 modified Metabolites + 5 x 10 3 Phenotype

microrna = mir 1993 two reports that a small (= micro) RNA modulates expression of a protein in the nematode C. elegans 2002 initial reports of a potential role of micrornas in cancer

mirna genes: Human 1,424 Mouse 720 Zebrafish 358 Berezikov Nat Rev Genetics 2011

micrornas microrna expression patterns in normal vs cancer tissues Colon Pros Brst ColonCa ProCa BrCa normal tissues cancer tissues Lu et al. Nature 435:834, 2005

microrna versus mrna expression patterns in normal human tissues ( 89 different epithelial tissues) Lu et al. Nature 435:834, 2005

2008

Nature Methods 2011

2007; Vol. 9:654-9

2011; Vol. 13:421-433 2007; Vol. 9:654-9

What is in a liquid biopsy? plasma buffy coat (leukocytes) erythrocytes, platelets cell-free: DNA microrna proteins metabolites Vietsch et al Pancreat Disord Ther 5:22; 2015

What is in a liquid biopsy? plasma buffy coat (leukocytes) erythrocytes, platelets cell-free: DNA microrna proteins metabolites + cancer cells cancer cells stroma Vietsch et al Pancreat Disord Ther 5:22; 2015

Detect metastatic disease in early stage cancer? plasma buffy coat (leukocytes) erythrocytes, platelets cell-free: DNA microrna proteins metabolites + cancer cells cancer cells stroma Vietsch et al Pancreat Disord Ther 5:22; 2015

serum before the initial surgery of stage I/II colon cancer Shivapurkar et al PLoS One 2014 e84686

Shivapurkar et al PLoS One 2014 e84686

The usual suspects approach... Shivapurkar et al PLoS One 2014 e84686

B Genome wide mir serum profile one patient with one without recurrent disease RC NRC hsa-mi-378 hsa-mir-92a hsa-mir-185 hsa-mir-1979 hsa-mir-25 hsa-mir-1977 hsa-mir-320a hsa-mir-1246 hsa-mir-320b hsa-mir-1974 hsa-mir-424 hsa-mir-374a hsa-mir-486-5p hsa-mir-223 hsa-mir-1274b hsa-mir-20b hsa-mir-126 hsa-mir-195 hsa-mir-451 hsa-mir-16 hsa-mir-103 hsa-mir-17 hsa-mir-140-3p hsa-mir-191 hsa-let-7c hsa-mir-15a hsa-mir-135a hsa-mir-1290 hsa-mir-1307 hsa-mir-30e hsa-mir-376a hsa-mir-26b hsa-mir-425 hsa-mir-29c hsa-mir-106b hsa-mir-107 hsa-mir-23b hsa-mir-494 hsa-mir-92b hsa-mir-192 hsa-mir-151-3p hsa-mir-320c hsa-mir-342-5p hsa-mir-122 hsa-mir-320d hsa-mir-30d hsa-mir-142-3p hsa-mir-150 hsa-mir-1975 hsa-mir-106a hsa-mir-19a hsa-mir-19b hsa-mir-374b hsa-mir-30a hsa-mir-135b hsa-mir-1274a hsa-mir-93 hsa-let-7b hsa-mir-15b hsa-mir-532-3p hsa-mir-221 hsa-mir-125a-5p hsa-mir-615-5p hsa-mir-1260 hsa-mir-596 hsa-mir-1287 hsa-mir-1915 hsa-mir-23a hsa-mir-21 hsa-mir-101 Shivapurkar et al PLoS One 2014 e84686

Shivapurkar et al PLoS One 2014 e84686

Monitor treatment response? plasma buffy coat (leukocytes) erythrocytes, platelets cell-free: DNA microrna proteins metabolites + cancer cells cancer cells stroma Vietsch et al Pancreat Disord Ther 5:22; 2015

LaConti et al. 2011 PLoS One e20687

Analysis of micrornas during PanCa progression ( Kras mutant model ) micrornas qrt-pcr animal model P48Cre x LSL/KrasG12D patients pancreatic cancer initiation and progression

Analysis of micrornas during PanCa progression ( Kras mutant model ) LaConti et al. 2011 PLoS One e20687

micrornas in the circulation & gemcitabine treatment mouse mutant K-ras Panc Ca model blood samples diagnosis profile treatment blood samples response profile micrornas

micrornas in the circulation & gemcitabine treatment mouse mutant K-ras Panc Ca model LaConti et al. 2011 PLoS One e20687

Shivapurkar et al. 2012 Int J Colorectal Dis

Circulating mir patterns in patients with pancreatic cancer treatment with capecitabine & lapatinib different times of treatment ( n=8 ) Tian, Shivapurkar et al. 2015 Oncology Letters in press

Evolutionary selection of cancer cell populations with distinct drivers of malignant progression

Evolutionary selection of cancer cell populations with distinct drivers of malignant progression from: Yates, Nature Rev Genetics 2012

Monitor the evolutionary selection of cancer cell populations / treatment response plasma buffy coat (leukocytes) erythrocytes, platelets cell-free: DNA microrna proteins metabolites + cancer cells cancer cells stroma Vietsch et al Pancreat Disord Ther 5:22; 2015

Food for Thought: Bad Tumor Markers Are Equal to Bad Drugs Daniel F. Hayes, MD U Mich, Ann Arbor

Ivana Peran Eveline Vietsch Anne Deslattes Joe LaConti Anna T Riegel Narayan Shivapurkar John Marshall Louis Weiner Claudine Isaacs Ruth A He Hartmut Juhl, Indivumed Hamburg, Germany Casper van Eijck, Erasmus U Rotterdam, Netherlands

Ivana Peran Eveline Vietsch