(Epi)Genetics in normal and malignant germ cell development. Leendert Looijenga, Department of Pathology, Lab. Exp. Patho-Oncol. Erasmus MC, Rotterdam OPTIMAL PATIENT CARE MM Basic and Transl. Oncol., October 23, 2017, 15.00-15.45 hours. Diagnosis -- Treatment -- Follow-up
Traditional classification(s) lack(s) biological basis. Extragonadal mainly type I & II Oosterhuis and Looijenga, Nat Rev Cancer 2005
Accepted WHO 2016 classification (testicular) II I III Spermatocytic seminoma
Human (sperm.) seminoma (tumor) [Type II-III] Morphology, epidemiology and behavior Stoop et al., Lab. Invest. 2001 mitosis/meiosis: XPA, SYCP1, SSX2-4 (Rajpert de Meyts et al., 2003: Chk2, p53, p16ink4d, MAGE-A4) Looijenga et al., Cancer Res 2006 acgh (>#9) DMRT1 (Doublesex and MAB-3 related transcription factor) Sem./Dysgerm Sperm. Sem.
Additional markers Sperm. Seminoma (Tumor) Animal model DMRT1
Recent paper NGS (story closed?)
GENERAL POPULATION OBSERVATIONS Focus Type II (TGCTs): Epidemiology. Historical (main) risk factors: - Cryptorchidism - Sub/infertility - Familial predisposition - Inguinal hernia - Birth weight - Hypospadias - DSD (intersex) TDS (THEORETICAL) ASSUMPTION: ALL (??????) Long term effect because of young patients & life expectancy: Infertility, Metabolic Syndrome, Vasc./Heart Damage, 2 nd Cancers
Precursor classification: acceptance WHO 2016. PRECURSOR TERMINOLOGY
Precursor classification: acceptance WHO 2016. PRECURSOR TERMINOLOGY
Overview pathology GCNIS related TGCT. COMMON PRECURSOR PRE-INVASIVE: Early gonadal development seminoma INTRATUBULAR: GCNIS (GB-DSD) 70% ~ 7 years Blocked PGC/gonocyte (EMBRYONIC GERM CELL) embr. carc. INVASIVE: PLURIPOTENT (TOTI-/OMNI-) seminoma embr.carc. germ cells PURE or MIXED yolk sac tum. teratoma choriocarc.
Overview pathology & biomarkers (serum); selection. PRE-INVASIVE: Early gonadal development OCT3/4 GCNIS (GB-DSD) COMMON PRECURSOR Blocked PGC/gonocyte (EMBRYONIC GERM CELL) seminoma embr. carc. INTRATUBULAR: OCT3/4 70% ~ 7 years OCT3/4 INVASIVE: PLURIPOTENT (TOTI-/OMNI-) OCT3/4 seminoma OCT3/4 embr.carc. Palumbo et al., 2001; Looijenga et al., 2003, De Jong et al., 2005, Cheng et al., 2006; De Jong and Looijenga, 2007, Van Casteren et al., 2008a,b, 2009, De Jong et al., 2008, Nonaka et al., 2009, Sonne et al, 2010, Gillis et al., 2011, Ushida et al., 2012, Eini et al., 2014, and many others. AFP yolk sac tum. teratoma germ cells hcg choriocarc.
Overview pathology & biomarkers (serum); selection. PRE-INVASIVE: Early gonadal development OCT3/4 SOX17 GCNIS (GB-DSD) COMMON PRECURSOR Blocked PGC/gonocyte (EMBRYONIC GERM CELL) seminoma embr. carc. INTRATUBULAR: OCT3/4 SOX17 70% ~ 7 years OCT3/4 SOX2 INVASIVE: PLURIPOTENT (TOTI-/OMNI-) OCT3/4 SOX17 seminoma OCT3/4 SOX2 embr.carc. SOX17 Palumbo et al., 2001; Looijenga et al., 2003, De Jong et al., 2005, Cheng et al., 2006; De Jong and Looijenga, 2007, Van Casteren et al., 2008a,b, 2009, De Jong et al., 2008, Nonaka et al., 2009, Sonne et al, 2010, Gillis et al., 2011, Ushida et al., 2012, Eini et al., 2014, and many others. AFP yolk sac tum. SOX2 teratoma germ cells hcg choriocarc.
SOX2 and 17 critical ES resp. PGC.
Biomarkers (WHO 2106) and dap (PGC/ES). Similar (protein) patterns normal development & (T)GCTs (highly informative diagnostic markers)
DSD & (T)GCT precursors (relevance pathologists). DSD guideline 2017: LEPO = referral center (NL/B)
Possible overdiagnosis GCNIS (delayed maturation). OCT3/4 and SCF (KITLG): Distinction between delayed matured and (pre-)gcnis -- versus ++ Stoop et al., 2008 delayed maturation: (OCT3/4+/ KITLG-) Sophia Children s Hospital: Start 2002= 12/160 (7.5%) = 2 (17%) progressed; Update: ~200 orchidopexy biopsies (till end 2013, >250 to date); pre-gcnis: 14 (7%); progression n=3 (21.4%; age after puberty), 1 bilateral in follow-up. Diagnosis based on (gonadal) tissue analysis; GWAS on TGCT cohorts Rapley et al., 2009; Kanetsky et al. 2009; Turnbull et al. 2010; Kratz et al., 2011; Ruark et al., 2013; Chung et al., 2013;.. + KITLG [OR = 2.69 highest to date] + SPRY4 [inh. MAPK, downstream KITLG] + BAK1 [downstream KITLG] + DMRT1 [sex determination] + TERT, ATFIP [telomere maintenance] + UCK2, HPGDS, CENPE, CLPTM1L, MAD1L1, RFWD3, TEX14, PPM1E
TECAC + SNP update TGCT (germline variations).
SNP and DSD (Hum. Reprod.: 2017 in press).
Novel susceptibility variants (SNPs) (not GWAS)
From germline genetics to (T)GCT epigenetics.
DNA methylation-based TGCT insights. CNV (heterogeneity) Genomic imprinting Pathway activity Lineage similarities
Conclusions so far and expectations. Optimal pathological (WHO)/clinical classification (also non-testicular) Identification of (general) risk population(s) ethnic origin (genetic basis) & clinical characteristics (TDS/DSD) (SNPs) Insight into pathogenesis (embryonic germ cell origin, including (epi)genetics) Identification (biology based) biomarkers (Tissue/Serum) OCT3-4/SOX2-17/(KITLG early age)/afp/hcg (Tissue/Serum)/... Risk stratification: * initial diagnosis * follow-up
TGCT versus ES/IPS cells clinical relevance Potential of malignant behavior = (T)GCT??
Daniela Salvatori, LUMC Pre-clinical in vivo model = bridge disciplines (?)
mrna profiling (cell lines vitro/vivo & TGCT)
mirna profiling (cell lines vitro/vivo) down
mirna profiling (cell lines vitro/vivo sera) EC TE (YST)
Overall conclusions and future perspectives Human germ cell tumors; a developmental disease. What we have: - Optimal pathological (WHO)/clinical classification (also non-testicular) - Identification of (general) risk population(s) ethnic origin (genetic basis) & clinical characteristics (TDS/DSD) (SNPs) - Insight into pathogenesis (embryonic germ cell origin) - Identification (biology based) biomarkers (Tissue/Serum) OCT3-4/SOX2-17/(KITLG early age)/afp/hcg (Tissue/Serum)/... What we will get: - Early stratification diagnosis (defined risk populations) - Understanding (genetic) heterogeneity (prim. tumor & metastases) - Informative molecular serum biomarkers (non-responders: relapse/resistance) - Putative target(s) treatment refractory disease
Rev. Fund, ESMO, ESPE, Deutsche KrebsHilfe, EORTC, Dr. Mildred Scheel Foundation Human germ cell tumors; a developmental disease. (missing KW, KB JWO, HS YvB) LEPO Urologists and Pathologists, Patients