Hypothesis/Strategy Identify genes downstream of AR that are important for survival of prostate cancer cells – AR signature genes – Regulators of AR signature Key drivers from Bayesian Genes with overlapping perturbation signatures Androgen ablation therapy will select for changes downstream of AR that maintain growth signal in absence of Androgens – Genes whose expression changes with disease progression – Genes subject to CNV with disease progression

Criteria for Prioritization AR signature Regulator of AR signature Evidence for mutation/CNV in cancer Evidence for expression changes/CNV changes with disease progression

2-stage Prioritization (Subjective) Weighting of criteria to create rank – AR sig / AR regulators / Mutation – Above + expression/CNV changes with grade Manual look at top 20 and select – Known genes removed – Priority for things which inhibition required – druggable

Literature: Implicated in colon cancer through CNA/Expression AS oligos block HBV propagation in HepG2 Mouse Gene Trap gets age-related pulmonary emphysema Validation Unclear whether need to inhibit or activate to reduce growth? Mechanism ?

Literature: Suppresses tumors of Hela in nude mice Binds c-Abl and MAPK1/ERK1 Adjacent to ASCL3 on chr 11 Disrupted in congenital Mental Retardation? Validation Activate Mechanism Tumor suppressor: Growth pathways

Literature: Phosphorylated by cell cycle kinase Aurora-C Modulates transcriptional activity of NRs (unliganded) Reduced TACC1 increases RARa & TRa activity Associated with endocrine therapy resistance in BRC Amplified in Breast Cancer (and Prostate Cancer) Validation Inhibit Mechanism NHR signaling

Literature: ACSL3 shows downregulation in high grade/met PRC Important for lipid biosynthesis/regulated by PPARd 5’ translocation partner of ETC1 in PRC Role in Lyn transport through golgi to PM KD reduces Lyn export Validation Inhibit? Triacsin C inhibitor available Mechanism Metabolism: lipid biosyn

Literature: Associated with Schizophrenia Associated with CNV Deleted in PRC Endocytic regulatory protein Regulator of axon elongation in neurons Validation Activate? Mechanism ?

Literature: Positive regulator of glycolytic function in glioblastomas Generates acetyl-CoA+OA from citrate and CoA Acetyl-CoA important for biosynthesis Regulated by AKT in lung carcinoma lines. siRNA resulted in growth arrest Validation Inhibit Hydroxycitrate inhibits Mechanism Metabolism: glycolysis/TCA

Literature: TSC2 is negative regulator of mTOR pathway. mTOR pathway activity associated with hormone- refractory PRC Validation Activator Fisetin inhibits mTOR pathway and activates TSC2 Mechanism Growth pathways

Literature: Global mediator of steroid receptor action in hormone- dep cancers FoxA1 in prostate cancer appears to act with AR in a combinatorial fashion to regulate transcription Low FoxA1 even with high AR predicts good prognosis In CPRC drives G1-S transition and G2/M transition Validation Inhibitor Mechanism NHR signaling

Literature: Inhibitor of Bax-induced apoptosis shown to interact with broad range of partners to inhibit many facets of apoptosis, such as ROS production, cytosolic acidification and calcium levels as well as endoplasmic reticulum stress signalling pathways activity is important in a large number of cancers, promoting metastasis by modulating actin dynamics Validation Inhibitor Mechanism Apoptosis

Literature: Mutations cause metabolic disease: 3-hydroxisobutyric aciduria Validation Not clear? Mechanism Metabolism: amino acid catabolism

Literature: Important for stromal survival/differentiation? Validation inhibit Mechanism Stromal regulation?

Validation Inhibit (labedipinedilol-A) Mechanism Apoptosis Literature Inhibition of human prostate cancer cells proliferation by a selective alpha1-adrenoceptor antagonist labedipinedilol-A involves cell cycle arrest and apoptosis.

Validation Inhibit (isoform) Mechanism epigenetic? Literature Upregulation associated with leukemias Downreg of isoform lacking transcriptional activator in CRC

Literature: Tumor suppressor? Region frequently lost in cancer; prostate, breast, HCC Secreted protein associated with stromal cells? Validation Activate Mechanism Stromal?

Metabolic Pathways IDH1 ACLY ACSL3 Beta-oxidation of lipids TCA Cycle

AR Signature Positively correlated gene network

AR Signature Negatively correlated gene network