Supplementary Table S1 . Summary of cell lines used.
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Cell line Purpose Reference H441 H441 responds to HGF in assays of cell migration; here, H441 is used as a functional MET agonist assay. Kong-Beltran et al. Cancer Cell 2004; 6: 75 H596 H596 proliferation increases upon HGF stimulation; here, H596 is used as a model of HGF-dependent MET activation. Kong-Beltran et al. Cancer Res 2006; 66: 283 U87MG The in vivo growth of U87MG is driven by an HGF/MET autocrine loop and the model has been used to investigate many anti-HGF and anti-MET therapeutics. Burgess et al. Cancer Res 2006; 66: 1721 DU145 DU145 cells scatter in response to HGF treatment; here, DU145 is used as a functional MET agonist assay. Humphrey et al. Am J Pathol 1995; 147: 386 HepG2 HepG2 responds to HGF in several functional assays including invasion and tubulogenesis ; here, HepG2 is used as a functional MET agonist assay. Neaud et al. Hepatology 1997; 26: 1458 Michaud et al. mAbs 2012; 4: 710 PHH (primary human hepatocytes) Hepatocytes have been shown to proliferation in response to HGF; here, human hepatocytes are used as a functional MET agonist assay. Nakamura et al. FEBS Letters 1987; 224:311 Gohda et al. JCI 1988; 81: 414 MKN45 MKN45 is MET amplified and MET is constitutively active and is used as a model of HGF-independent MET activation. Smolen et al. PNAS 2006; 103: 2316 H1437 H1437 has an R988C juxtamembrane domain MET mutation and was used to determine if mutant MET can be internalized. Ma et al. Cancer Res 2005; 65:1479 NIH3T3-M1149T The M1149T MET tyrosine kinase domain mutation was introduced into NIH3T3 to determine if mutant MET can be internalized. Schmidt et al. Oncogene 1999; 18:2343 LoVo LoVo expresses unprocessed 190kD pro-MET and was used to determine if this MET alteration can be internalized. Mondino et al. Mol Cell Biol 1991; 11: 6084 EBC-1 EBC-1 is MET amplified and MET is constitutively active and is used as a model of HGF-independent MET activation. Lutterbach et al. Cancer Res 2007; 67:2081 ementary Table S1. Summary of cell lines used.
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Supplementary Table S1 . Summary of cell lines used. - PowerPoint PPT Presentation
Transcript of Supplementary Table S1 . Summary of cell lines used.
Cell line Purpose Reference
H441 H441 responds to HGF in assays of cell migration; here, H441 is used as a functional MET agonist assay.
Kong-Beltran et al. Cancer Cell 2004; 6: 75
H596 H596 proliferation increases upon HGF stimulation; here, H596 is used as a model of HGF-dependent MET activation.
Kong-Beltran et al. Cancer Res 2006; 66: 283
U87MG The in vivo growth of U87MG is driven by an HGF/MET autocrine loop and the model has been used to investigate many anti-HGF and anti-MET therapeutics.
Burgess et al. Cancer Res 2006; 66: 1721
DU145 DU145 cells scatter in response to HGF treatment; here, DU145 is used as a functional MET agonist assay.
Humphrey et al. Am J Pathol 1995; 147: 386
HepG2 HepG2 responds to HGF in several functional assays including invasion and tubulogenesis ; here, HepG2 is used as a functional MET agonist assay.
Neaud et al. Hepatology 1997; 26: 1458Michaud et al. mAbs 2012; 4: 710
PHH (primary human hepatocytes)
Hepatocytes have been shown to proliferation in response to HGF; here, human hepatocytes are used as a functional MET agonist assay.
Nakamura et al. FEBS Letters 1987; 224:311Gohda et al. JCI 1988; 81: 414
MKN45 MKN45 is MET amplified and MET is constitutively active and is used as a model of HGF-independent MET activation.
Smolen et al. PNAS 2006; 103: 2316
H1437 H1437 has an R988C juxtamembrane domain MET mutation and was used to determine if mutant MET can be internalized.
Ma et al. Cancer Res 2005; 65:1479
NIH3T3-M1149T The M1149T MET tyrosine kinase domain mutation was introduced into NIH3T3 to determine if mutant MET can be internalized.
Schmidt et al. Oncogene 1999; 18:2343
LoVo LoVo expresses unprocessed 190kD pro-MET and was used to determine if this MET alteration can be internalized.
Mondino et al. Mol Cell Biol 1991; 11: 6084
EBC-1 EBC-1 is MET amplified and MET is constitutively active and is used as a model of HGF-independent MET activation.
Lutterbach et al. Cancer Res 2007; 67:2081
SNU-5 SNU-5 is MET amplified and MET is constitutively active and is used as a model of HGF-independent MET activation.
Smolen et al. PNAS 2006; 103: 2316
Caki-1 Caki-1 proliferates in response to HGF and HGF protects Caki-1 cells from apoptosis; here Caki-1 is used as a functional MET agonist assay.
Horie et al. J Urology 1999; 161: 990Huang et al. Tumor Biol 2014; doi10.1007/s13277-014-1774-7
Supplementary Table S1. Summary of cell lines used.
Supplementary Table S2. Summary of Repeat-Dose Toxicity Study with LY2875358
Species Strain
No./Sex/Group Age
Doses, Route,
Duration
Parameters Evaluated
Noteworthy Findings
Monkey, Cynomolgus
3 (0.3, 7 mg/kg) 6 (0, 180 mg/kg)
1.5-2.5 years
0a, 0.3, 7, and 180 mg/kg Q7D
Intravenous
5 wk + 16 wk recovery
phase
TKb, survival, BW, FC, clin signs, phys, ophthal, ECG, safety pharmc, ADA, PDd, clin pathe, pathf, urine biomarkersg, IHCh
≥0.3 mg/kg: thyroid weight, follicular thyroid dilatation, ↑ECD 7 mg/kg (M): Enlarged thyroid 180 mg/kg (F): Blood in urine - Weeks 2 and 5
Recovery Phase: Limited to detection of ADAs in 3 of 12 animalsi NOAEL = 180 mg/kg
Abbreviations: = increase; ADA = antidrug antibody; BW = body weight; clin = clinical; CNS = central nervous system; ECD = extracellular domain; ECG = electrocardiography; ELISA = enzyme-linked immunosorbent assay; FC = food consumption; F = female; IgG = immunoglobulin G; IHC = immunohistochemistry; M = male; MET = mesenchymal-epithelial transition factor; NOAEL = no-observed-adverse-effect level; No. = Number; ophthal = ophthalmic examinations; path = pathology; PD = pharmacodynamics; phys = physical examinations; safety pharm = safety pharmacology; TK = toxicokinetics; wk = week; Q7D = Once every 7 days
a Vehicle control group. Vehicle = 10 mM sodium citrate buffer (pH 6.0), 150 mM sodium chloride, 0.02% polysorbate 80 in sterile water for injection.
b Total IgG LY2875358 assay; extracellular domain of MET analysis. c Respiratory exams, neurologic exams with detailed CNS/behavioral observations, body temperature.
d Total MET analysis in kidney, skin, and liver by ELISA and Western analysis. e Hematology, clinical chemistry, and urinalysis. f Organ weights, gross pathology, and histopathology. g Neutrophil gelatinase-associated lipocalin (NGAL), microalbumin, clusterin. h Ki67 staining in liver, kidney, and spleen. i Presence of drug antibody may have interfered with assay’s ability to detect ADAs at the end of the recovery
phase as well as at the end of the treatment phase.
