The Texas Pediatric PDX Core Peter J. Houghton, Ph.D., Director Greehey Children s Cancer Research Institute UT Health Science Center San Antonio
GOALS To successfully heterograft and propagate 200-250 new Patient Derived Xenograft (PDX) preclinical models from Hispanic and underserved children in Texas. To molecularly characterize each model and provide annotated deidentified patient demographic data associated with each tumor model. To establish a database web portal to allow Core users to access patient demographic, genomic and other data associated with individual models. To provide well-characterized, annotated, cryo-preserved xenograft tissues to researchers in Texas and beyond. To provide a Training Course for interested researchers interested in developing and using PDX models in research. Develop a novel zebrafish xenograft bioassay. In years 3-5, we will develop and validate a novel metastasis bioassay using the zebrafish model system.
ORGANIZATION De-identified Patient Database UTHSCSA -Tomlinson UHS/Methodist Sample acquisition Blood/Tumor/Shipping UTSWMC Skapek Sample acquisition Blood/Tumor/Shipping TPPDX Core Houghton/Kurmasheva Heterografting Coding Website Cryo-preservation Characterization Distribution Committee Exome sequence RNASeq Immunophenotype (leukemias) Research Laboratories
Leukemia PDX Models Established Specimen ID/Source Gende r Race/Ethnicity Age Diagnosis Relapse Y/N Germline DNA, Y/N/P 0453-000, UT Health/UHS M White/Hispanic 16 years Pre B ALL N Y 0485-000, UT Health/UHS F White/Hispanic 25 years Pre BALL N Y 0487-000, UT Health/UHS F White/Hispanic 7 y 2 m B Cell, ALL Y (2nd) Y 0495-000, UT Health/UHS M White/Hispanic 19 y 8 m Pre B ALL N N 0426-000, UT Health/UHS F White/Hispanic P AML, secondary to treatment for Ewing's sarcoma N N 0501-000, UT Health/UHS M White/Hispanic 2 y 1 m B Cell, ALL N Y MCH 0026, Methodist CH M White/Hispanic 2 years Pre B ALL N N MCH 0027, Methodist CH M White/Hispanic 19 y 10 m T- cell ALL Y N MCH 0029, Methodist CH F White/Not Hispanic 27 y 3 m AML N N MCH 0030, Methodist CH F Not White/Not Hispanic 13 y 5 m ALL N N MCH 0031, Methodist CH F White/Hispanic 7 m ALL N N MCH 0032, Methodist CH M White/Not Hispanic 8 y ALL N N Cryopreserved 1962 vials at 10 7 cells/vial
Solid Tumor PDX Models Established Specimen ID/Source Gender Race/ Ethnicity Age Diagnosis Relapse Y/N Primary/ Metastatic Germline DNA, Y/N/P 0462-000 UY Health/UHS F Hispanic 6 Medulloblastoma N P Y 0466-000 UY Health/UHS F NH White 2 Medulloblastoma N P Y 0494-000 UY Health/UHS M NH White 2 Hepatoblastoma N P Y CPRIT 1739 UTSW-Dallas M NH White 3 Hepatoblastoma N P Y CPRIT 1748 UTSW-Dallas F NH White 11 mo Hepatoblastoma N P Y CPRIT 1763 UTSW-Dallas F AA 2 Wilms N P Y CPRIT 1796 UTSW-Dallas F AA CPRIT 1795 UTSW-Dallas M Hispanic 16 17 Ovarian N P Y Testicular N P Y COG851210 St. Mary s, FL P Germ Cell P 14 samples < 3months, pending
Samples Viably Cryopreserved for Future Distribution Leukemias Specimen ID/Source Diagnosis Current Passage Vials Frozen (passage) 0453-000, UT Health/UHS Pre B ALL 3 F1-16 F2-42 F1-30 0485-000, UT Health/UHS Pre BALL 4 F2-254 F3-37 0487-000, UT Health/UHS B Cell, ALL 2 F1-84 F1-322 0495-000, UT Health/UHS Pre B ALL 4 F2 537 F3-324 0501-000, UT Health/UHS B Cell, ALL 2 F1-43 MCH 0026, Methodist CH Pre B ALL 3 F1-220 F2-21 MCH 0027, Methodist CH T- cell ALL 1 F1-5 MCH 0030, Methodist CH ALL 1 F1-6 MCH 0031, Methodist CH ALL 1 F1-21 10 7 cells/vial
Samples Viably Cryopreserved for Future Distribution Solid Tumors Specimen ID Source Gender Race/Ethnicity Age Diagnosis Relapse Primary/ Germline DNA Y/N Metastatic Y/N/P 1. 0462-000 UT Health/UHS F Hispanic 6 Medulloblastoma N P Y 2. CPRIT 1753 UT Southwestern M AA 11 Wilms N P Y 3. 0466-000 UT Health/UHS F NH White 2 Medulloblastoma N P Y 4. CPRIT 1763 UT Southwestern F AA 2 Wilms N P Y 5. 0500-000 UT Health/UHS M Hispanic 14 Osteosarcoma N P N 6. CPRIT 1796 UT Southwestern F AA 16 Ovarian ca, GCT N P Y 7. CPRIT 054 UT Southwestern M Hispanic 10 Refractory ERMS Y P 8. CPRIT 1739 UT Southwestern M NH White 3 Hepatoblastoma N P Y 9. 0471-2 UT Health/UHS M Hispanic 13 Osteosarcoma N M Y 10. 0494-000 UT Health/UHS M NH White 2 Hepatoblastoma N P Y 11. CPRIT 0029 UT Southwestern F AA 4 Wims- Pulmonary Met Y M 12. CPRIT 1795 UT Southwestern M Hispanic 17 Testicular-Mixed GCT N P Y 13. CPRIT 1837 UT Southwestern M NH White 15 Hodgkins Lymphoma 14. CPRIT 1853 UT Southwestern Neuroblastoma 6 pieces/vial
Why PDX Models of Childhood Cancer? They maintain the fidelity (pathology, genetics and epigenetics) of childhood cancers. Expanded panels of PDX models will simulate clinical heterogeneity more accurately. PDX models are valuable in elucidating biology and therapy of childhood cancer. Models have been valuable in developing effective therapies with lower toxicity.
PPTP Data: Activity of Standard and Novel Cytotoxic Agents Red = Maintained Complete Regressions Orange = Complete Regression Yellow = Partial Regression (>50%)
Comparison of Eribulin Pharmacokinetics in Mouse and Adult Patients 10000 Mouse 10000 Human Eribulin plasma conc. (nm) 1000 100 10 1 1 mg/kg Eribulin plasma conc. (nm) 1000 100 10 1 0.1 0.17 5 10 15 20 Hours Kolb EA et al Ped Blood Cancer 2013 0.1 0 24 48 72 96 120 144 168 Hours Hours Morgan RJ et al CCP 2015
ES-4 Ewing Sarcoma Xenograft: Greater than additive activity for Eribulin + Irinotecan 10.0 Tumor Volume cm3 1.0 0.1 Control 0 1 2 3 4 5 6 7 8 9 1011121314151617181920 Time (Weeks) 10.0 Tumor Volume cm3 1.0 0.1 Eribulin R x 0 1 2 3 4 5 6 7 8 9 1011121314151617181920 Time (Weeks) 10.0 Tumor Volume cm3 1.0 0.1 Irinotecan R x 0 1 2 3 4 5 6 7 8 9 1011121314151617181920 Time (Weeks) 10.0 Tumor Volume cm3 1.0 0.1 Vincristine (1 mg/kg) R x 0 1 2 3 4 5 6 7 8 9 1011121314151617181920 10.0 Tumor Volume cm3 1.0 0.1 Eribulin + Irinotecan R x 0 1 2 3 4 5 6 7 8 9 1011121314151617181920 10.0 Tumor Volume cm3 1.0 0.1 Vincristine + Irinotecan R x 0 1 2 3 4 5 6 7 8 9 1011121314151617181920 Time (Weeks) Time (Weeks) Time (Weeks)
Summary of Median Event-Free Survival for Sarcoma models Treated with Eribulin Or Vincristine Combined with Irinotecan Xenograft Model Tumor Type Eribulin + Irinotecan Vincristine + Irinotecan P EFS (days) Rh10 Alveolar rhabdomyosarcoma >91 59.3 <0.001 Rh30 Alveolar rhabdomyosarcoma 90.8 >160 <0.001 Rh41 Alveolar rhabdomyosarcoma 80.2 79.2 0.735 Rh65 Alveolar rhabdomyosarcoma >133 121.1 0.011 ES-4 Ewing sarcoma 62.9 35.7 <0.001 CHLA258 Ewing sarcoma 81.6 35.7 <0.001 SKNEP1 Ewing sarcoma >133 >133 1.000 KT-11 Wilms >77 29.7 <0.001 KT-13 Wilms 74.7 77.9 0.312 RBD1 Rhabdoid >96.1 >96.1 1.000 KT-14 Rhabdoid 96.4 56.3 0.004 Eribulin 1 mg/kg IP days 1,8 repeated at day 21 Vincristine 1.0 mg/kg IP weekly x 6 (single agent) Vincristine 0.5 mg/kg IP weekly x 6 (combination) Irinotecan 2.5 mg/kg IP days 1-5, repeated at day 21.
MEK Inhibitors in Childhood Glioma Low-grade glioma most common brain tumor in children Therapy is limited in those with unresectable disease BRAF aberrations in 70-90% of JPAs, most commonly BRAF-KIAA1549 fusion protein Alternate BRAF aberrations described including BRAFV 600E mutation in 10-15% of JPA
Only Mutant BRAF Glioma Respondes to MEK Inhibition CONTROL BT-35 WT BRAF CONTROL BT-40 V600E
Phase I Responses to Selumetinib in Pediatric Low-Grade Glioma (PBTC 029 Trial)
Inhibition of MEK Suppresses TORC1 in BT-40 BRAF-mutant Tumors BT-35 BT-40 Control 4/0 4/2 Control 4/0 4/2 perk1/2 ERK1/2 ps6 S6 p4e-bp1 4E-BP1 pakt Akt GAPDH
Regulation of TORC1 by DNA Damage is Complex Fig5h.' ATM' DNA'damage' Growth' Signals' p53' p63' DNA.PK' Sestrin1/2' REDD1' Akt' AMPK' Raf1'(c.Raf)' TSC1/TSC2' MEK'1/2' p70s6k' 4E.BP1' mtorc1' p70s6k P' ERK'1/2' 4E.BP1 P' rs6' rs6 P' Cam et al JBC, 2015
BT40 (Passage 10) Scramble TSC2 shrna Scramble TSC2 shrna 0 2 5 0 2 5 0 2 5 0 2 5 Tramatinib (nm) TSC2 p-erk1/2 ERK1/2 p-s6k p-s6 p-4ebp1 4EBP1 GAPDH 12h 24h
In BT-40 Cells BRAF Regulates TORC1 via TSC Inhibition 100 Tramatinib (nm) DMSO Scramble shrna 1 2 5 10 20 10 TSC2 shrna myrakt1 1 myrakt3 Scramble TSC2-shRNA myrakt1 myrakt3 0.1 0 5 10 15 20 25
BT40 Trametinib+ Rapamycin Study R x R x Regrowth delay R x
Goals for Next Year Increase the acquisition of tumor samples, UTSW (brain tumors & leukemias), MD Anderson affiliation for samples. Start molecular characterization (exome sequence, RNAseq). Develop the website to advertise PDX models for Research. Develop the Zebrafish model for primary transplantation.
Texas Pediatric PDX Core UTHealth San Antonio Peter Houghton Gail Tomlinson Raushan Kurmasheva Myron Ignatius Alison Grimes Assanasen Chatchawin Abhik Bandyopadhyay Anna Rogojina Luz Perez Prado UTSW Dallas Stephen Skapek James Armatruda Dinesh Rakheja Bo Yao Jiwoon Kim Cristyn Bransteter Erin Butler