Supplementary Table 1 - PDF file 282K, Supplemental table listing primer sequences utilized throughout study

PDF file - 410K, Dose responses, transcriptional, biological, biochemical, and in vivo supplemental data

<p>Supplemental Methods and Material</p>

Supplementary Figure 1,2 - PDF file 56K, Supplemental data demonstrating that scheduling of IR and ADT treatment does not alter cell survival and DNA damage induces AR mediated gene transcription in a dose dependent fashion

Supplementary Figure 7 - PDF file 79K, Supplemental data further demonstrating that androgen deprivation sensitizes CRPC cells to IR

Supplementary Figure 3,4 - PDF file 443K, Supplemental data demonstrating that exposure to IR promotes cell cycle arrest and AR activity regulates genes required for DNA damage repair

Abstract RB loss occurs commonly in neoplasia but its contributions to advanced cancer have not been assessed directly. Here we show that RB loss in multiple murine models of cancer produces a prometastatic phenotype. Gene expression analyses showed that regulation of the cell motility receptor RHAMM by the RB/E2F pathway was critical for epithelial–mesenchymal transition, motility, and invasion by cancer cells. Genetic modulation or pharmacologic inhibition of RHAMM activity was sufficient and necessary for metastatic phenotypes induced by RB loss in prostate cancer. Mechanistic studies in this setting established that RHAMM stabilized F-actin polymerization by controlling ROCK signaling. Collectively, our findings show how RB loss drives metastatic capacity and highlight RHAMM as a candidate therapeutic target for treating advanced prostate cancer. Cancer Res; 77(4); 982–95. ©2016 AACR.

<p>Supplemental Figure S1. RB loss promotes cell migration and invasion. Supplemental Figure S2. RB regulates RHAMM expression. Supplemental Figure S3. RB/E2F Binds on the RHAMM Promoter. Supplemental Figure S4. RHAMM overexpression drives cancer cell migration, invasion, and EMT. Supplemental Figure S5. RHAMM Inhibition Diminishes Migration and Invasion, but not Cellular Proliferation. Supplemental Figure S3. RB/E2F Binds on the RHAMM Promoter.</p>

PDF file - 91K

<p>PDF file - 50K, Primer sequences used in manuscript.</p>

in silico docking information

Supplementary Methods - PDF file 79K, Additional materials and methods not described in main manuscript

<p>PDF file - 176K, Supplementary Figure S1: USP22 Specifically Promotes AR Recruitment to Target Loci. Supplementary Figure 2: Depletion of USP22 can be mediated by multiple sequences.Supplementary Figure S3: AR Ubiquitylation Levels are not Altered in Response to USP22 Depletion.</p>

Supplementary Figure Legends - PDF file 66K, Figure legends corresponding to supplemental figures

<p>Supplementary Figure Legends Supplemental Figure S1. RB loss promotes cell migration and invasion. Supplemental Figure S2. RB regulates RHAMM expression. Supplemental Figure S3. RB/E2F Binds on the RHAMM Promoter. Supplemental Figure S4. RHAMM overexpression drives cancer cell migration, invasion, and EMT. Supplemental Figure S5. RHAMM Inhibition Diminishes Migration and Invasion, but not Cellular Proliferation.</p>

Supplementary Figure 5,6 - PDF file 143K, Supplemental data demonstrating that DNAPKcs knockdown is maintained in growth studies and ATM expression and activity is regulated by DNAPKcs knockdown and inhibition

Supplementary Figure 8 - PDF file 140K, Supplemental data demonstrating that DNAPKcs is required for DNA repair in the presence of androgen

The assessment of human health hazards posed by chemicals traditionally relies on toxicity studies in experimental animals. However, most chemicals currently in commerce do not meet the minimum data requirements for hazard identification and dose-response analysis in human health risk assessment. Previously, we introduced a read-across framework designed to address data gaps for screening-level assessment of chemicals with insufficient in vivo toxicity information (Wang et al., 2012). It relies on inference by analogy from suitably tested source analogues to a target chemical, based on structural, toxicokinetic, and toxicodynamic similarity. This approach has been used for dose-response assessment of data-poor chemicals relevant to the U.S. EPA's Superfund program. We present herein, case studies of the application of this framework, highlighting specific examples of the use of biological similarity for chemical grouping and quantitative read-across. Based on practical knowledge and technological advances in the fields of read-across and predictive toxicology, we propose a revised framework. It includes important considerations for problem formulation, systematic review, target chemical analysis, analogue identification, analogue evaluation, and incorporation of new approach methods. This work emphasizes the integration of systematic methods and alternative toxicity testing data and tools in chemical risk assessment to inform regulatory decision-making.