Short-term Progression-free Survival Research Articles

Alterations of the bile microbiome is associated with progression-free survival in pancreatic ductal adenocarcinoma patients

BackgroundPatients with pancreatic ductal adenocarcinoma (PDAC) display an altered oral, gastrointestinal, and intra-pancreatic microbiome compared to healthy individuals. However, knowledge regarding the bile microbiome and its potential impact on progression-free survival in PDACs remains limited.MethodsPatients with PDAC (n = 45), including 20 matched pairs before and after surgery, and benign controls (n = 16) were included prospectively. The characteristics of the microbiomes of the total 81 bile were revealed by 16 S-rRNA gene sequencing. PDAC patients were divided into distinct groups based on tumor marker levels, disease staging, before and after surgery, as well as progression free survival (PFS) for further analysis. Disease diagnostic model was formulated utilizing the random forest algorithm.ResultsPDAC patients harbor a unique and diverse bile microbiome (PCoA, weighted Unifrac, p = 0.038), and the increasing microbial diversity is correlated with dysbiosis according to key microbes and microbial functions. Aliihoeflea emerged as the genus displaying the most significant alteration among two groups (p < 0.01). Significant differences were found in beta diversity of the bile microbiome between long-term PFS and short-term PFS groups (PCoA, weighted Unifrac, p = 0.005). Bacillota and Actinomycetota were identified as altered phylum between two groups associated with progression-free survival in all PDAC patients. Additionally, we identified three biomarkers as the most suitable set for the random forest model, which indicated a significantly elevated likelihood of disease occurrence in the PDAC group (p < 0.0001). The area under the receiver operating characteristic (ROC) curve reached 80.8% with a 95% confidence interval ranging from 55.0 to 100%. Due to the scarcity of bile samples, we were unable to conduct further external verification.ConclusionPDAC is characterized by an altered microbiome of bile ducts. Biliary dysbiosis is linked with progression-free survival in all PDACs. This study revealed the alteration of the bile microbiome in PDACs and successfully developed a diagnostic model for PDAC.

Pretreatment MR-based radiomics in patients with glioblastoma: A systematic review and meta-analysis of prognostic endpoints

PurposeRecent studies have shown promise of MR-based radiomics in predicting the survival of patients with untreated glioblastoma. This study aimed to comprehensively collate evidence to assess the prognostic value of radiomics in glioblastoma. MethodsPubMed-MEDLINE, Embase, and Web of Science were searched to find original articles investigating the prognostic value of MR-based radiomics in glioblastoma published up to July 14, 2023. Concordance indexes (C-indexes) and Cox proportional hazards ratios (HRs) of overall survival (OS) and progression-free survival (PFS) were pooled via random-effects modeling. For studies aimed at classifying long-term and short-term PFS, a hierarchical regression model was used to calculate pooled sensitivity and specificity. Between-study heterogeneity was assessed using the Higgin inconsistency index (I2). Subgroup regression analysis was performed to find potential factors contributing to heterogeneity. Publication bias was assessed via funnel plots and the Egger test. ResultsAmong 1371 abstracts, 18 and 17 studies were included for qualitative and quantitative data synthesis, respectively. Respective pooled C-indexes and HRs for OS were 0.65 (95 % confidence interval [CI], 0.58–0.72) and 2.88 (95 % CI, 2.28–3.64), whereas those for PFS were 0.61 (95 % CI, 0.55–0.66) and 2.78 (95 % CI, 1.91–4.03). Among 4 studies that predicted short-term PFS, the pooled sensitivity and specificity were 0.77 (95 % CI, 0.58–0.89) and 0.60 (95 % CI, 0.45–0.73), respectively. There was a substantial between-study heterogeneity among studies with the survival endpoint of OS C-index (n = 9, I2 = 83.8 %). Publication bias was not observed overall. ConclusionPretreatment MR-based radiomics provided modest prognostic value in both OS and PFS in patients with glioblastoma.

Evaluation of progression-free survival as a surrogate endpoint in primary CNS lymphoma: a systematic review and meta-analysis.

Purpose: To evaluate progression-free survival (PFS) as early surrogate endpoints for overall survival (OS) in primary CNS lymphoma (PCNSL). Methods: PubMed, Embase and Cochrane Central Library were searched up to 7June 2022. Trial-level analyses were performed by weighted linear regression of logarithmic hazard ratios for PFS and OS. Treatment arm-level analyses were performed between PFS rates and 3- or 5-year OS rates. Results: 1471 PCNSL patients in ninerandomized control trials were included. PFS was associated with OS (r=0.750; 95% CI: 0.228-0.937). Strong linear correlations existed between 1-, 2- and3-year PFS and 3-year OS (r =0.896-0.928), moderate or weak correlations existed between 3- to 6-month PFS and 3-year OS, 3-month to 5-year PFS and 5-year OS. Conclusion: Short-term PFS can validly substitute forlong-term OS in PCNSL.

Nivolumab and AVD in Early-Stage Unfavorable Hodgkin Lymphoma: Follow-up Analysis of the Randomized GHSG Phase II Nivahl Trial

Background Anti-PD1 inhibition is increasingly investigated as first-line treatment of classical Hodgkin lymphoma (HL), usually either in sequential or concomitant combination with other therapeutics. While impressive response rates and short-term progression-free survival are observed, long-term efficacy and safety as well as quality-of-life (QoL) remain unknown. Herein we present the preplanned 3-year follow-up (FU) analysis of the GHSG phase II NIVAHL trial in early-stage unfavorable HL. Methods NIVAHL is an investigator-sponsored randomized phase II trial (NCT03004833) financially supported by Bristol-Myers Squibb which was conducted at 28 GHSG trial centers and enrolled 109 patients aged 18-60 years with centrally confirmed first diagnosis of early-stage unfavorable HL. Patients received either fully concomitant (4xnivo-AVD; arm A) or sequential (4xnivolumab, 2xnivo-AVD, 2xAVD; arm B) nivolumab-based 1st-line treatment, each followed by 30Gy involved-site radiotherapy (IS-RT). The primary analysis showed feasibility with a favorable safety profile and outstanding 1-year efficacy (Bröckelmann JAMA Oncol 2020). During FU, patients were monitored regularly for signs of HL relapse, persisting or late toxicities and QoL by the EORTC QLQ-C30 questionnaire. This FU analysis of key secondary endpoints including progression-free (PFS) and overall survival (OS), long-term safety and QoL was preplanned at three years after the registration of the last patient. Results With a median FU of 40 months, no PFS or OS events were observed since the NIVAHL primary analysis. Importantly, no patient with a partial remission by central response assessment required further treatment and all 7 patients converted to a complete remission during FU. With no deaths recorded, 3-year OS is 100% in both arms and 3-year PFS estimates are 98% and 100% in arm A and B, respectively (Figure 1). No second primary malignancies were reported and overall, no new safety signals were identified: While any treatment for potentially treatment-associated morbidities was required in 13% of patients, no patient required long-term corticosteroid treatment at last FU. Mean forced expiratory pressure in one second (FEV1) at baseline and 3 years after enrollment was 91.1% (standard deviation (SD) 13.6%) and 94.7% (SD 13%), respectively. Mean diffusion capacity for carbon monoxide adjusted for hemoglobin (DLCOc) at baseline and 3 years after enrollment was 86.2% (SD 16.5%) and 81.0% (SD 22.6%), respectively. No cardiac events >°1 were reported during FU and left ventricular ejection fraction (LVEF) was in the normal range in 93% of patients at last FU. The median global health score and global QoL by EORTC QLQ-C30 improved over time and were 64 (SD 22) and 80 (SD 16) at baseline, 76 (SD 17) and 87 (SD 15) 1 year after enrollment and 82 (SD 15) and 92 (SD 11) 3 years after enrollment, respectively. Relevant fatigue existed prior to treatment (mean fatigue score difference from age- and sex-adjusted German population reference value: 22 (SD 30)), briefly worsened on-treatment (after chemotherapy: 38 (SD 27), after radiotherapy: 27 (SD 23)) but resolved over time (1 year after enrollment: 10 (SD23), 3 years after enrollment: -3 (SD 18); Figure 2). Conclusion This preplanned FU analysis of the largest anti-PD1 HL first-line trial to date confirms the outstanding efficacy and favorable safety profile of this therapeutic approach. Additionally, QoL and fatigue appear relevantly improved compared to baseline during prolonged FU. Building on NIVAHL, the upcoming GHSG phase II INDIE trial (NCT04837859) will investigate an individualized immunotherapy with tislelizumab in early-stage unfavorable HL patients. Since likely not all patients require the full course of chemoimmunotherapy and consolidative IS-RT, INDIE will be the first trial investigating a chemo- and radiotherapy-free 1st-line HL treatment in optimally responding patients. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Prognostic Significance of Clinical Factors Including BRCA Mutation in Epithelial Ovarian, Peritoneal, Fallopian Tube Cancer.

We analyzed the survival outcomes of patients with epithelial ovarian, peritoneal, or fallopian tube cancer with BRCA1/2 mutations and the clinical factors associated with the prognosis of these cancers. We included patients who had been diagnosed with and treated for epithelial ovarian, peritoneal, or fallopian tube cancer and had undergone germline BRCA testing in six hospitals between January 2012 and December 2019. Of the 378 identified patients, 76 (20.1%) carried a BRCA1/2 mutation. Progression-free survival (PFS) and overall survival (OS) did not differ between patients with and without BRCA1/2 mutation. Multivariate analysis for 18 months after the primary treatment showed higher PFS in the BRCA1/2 mutation group (p=0.024). Subgroup analysis in patients with high-grade serous carcinoma showed that BRCA1/2 mutation was an independent favorable prognostic factor for PFS (p=0.035). Subgroup analysis of patients with stage III or IV disease demonstrated an independent gain in PFS in patients with BRCA1/2 mutation (p=0.015). Neoadjuvant chemotherapy as a primary treatment was related to poor PFS (p<0.001) and reduced OS (p=0.005). Having a germline BRCA1/2 mutation improved short-term PFS in patients with epithelial ovarian, peritoneal, or fallopian tube cancer. Elevated initial CA125 level and primary neoadjuvant chemotherapy were related to poor prognosis.

Clinical and molecular characteristics of patients with short- and long-term progression-free survival in the phase IIIb OPINION study of maintenance olaparib for patients with non-germline BRCA1/BRCA2-mutated platinum-sensitive relapsed ovarian cancer (306)

<b>Objectives:</b> The OPINION study (NCT03402841) demonstrated the activity of maintenance olaparib in patients (pts) with platinum-sensitive relapsed ovarian cancer (PSR OC) without a germline <i>BRCA1</i>/<i>BRCA2</i> mutation (non-g<i>BRCA</i>m), with an overall median progression-free survival (PFS) of 9.2 months (mo) (Poveda <i>et al</i>. ASCO 2021). We further explored clinical and molecular characteristics in pts with long-term (LT) and short-term (ST) PFS. <b>Methods:</b> Pts with non-g<i>BRCA</i>m PSR OC and ≥2 prior lines of platinum-based chemotherapy (PBC) incomplete/partial response (CR/PR) to their last PBC received maintenance olaparib (300 mg bid) until progression/unacceptable toxicity. Here, pts were grouped by time from the first dose until disease progression (RECIST v1.1) or death: >18 mo for the LT group and <4 mo for the ST group (chosen based on radiologic scan timings and follow up). Pts who discontinued the study >18 mo after the first dose were also in the LT group. Samples were analyzed using the Myriad myChoice HRD plus and BRACAnalysis CDx assays. Molecular analyses included: tumor <i>BRCA</i>m (t<i>BRCA</i>m); somatic <i>BRCA</i>m (s<i>BRCA</i>m), defined as the presence of a t<i>BRCA</i>m and no g<i>BRCA</i>m; homologous recombination deficiency (HRD), defined as a genomic instability score ≥42 and/or t<i>BRCA</i>m; homologous recombination repair (HRR) gene mutations, excluding <i>BRCA</i>m (non<i>-BRCA</i> HRRm), based on a prespecified 13-gene panel; and <i>TP53</i> mutation status. <b>Results:</b> Of 279 pts in OPINION, 67 (24%) and 65 (23%) had LT and ST PFS, respectively. At diagnosis, 45/67 pts (67%) in the LT group had FIGO stage III and 15 (22%) had stage IV disease; 47/65 (72%) in the ST group had stage III and 12 (18%) had stage IV disease. Median (range) prior PBC regimens was 2 (2-5) and 2 (2-6) in the LT and ST groups, respectively. About 34/67 (51%) and 12/65 (18%) pts achieved CR to their last PBC, and 33/67 (49%) and 52/65 (80%) had PR in the LT and ST groups, respectively; 48/67 (72%) and 42/65 (65%) had a platinum-free interval of ≥12 mo in the LT and ST groups, respectively. Fifteen out of 67 (22%) and 5/65 pts (8%) were found to have a t<i>BRCA</i>m in the LT and ST groups, respectively; 47/67 (70%) and 25/65 pts (40%) had HRD-positive tumors. Fifteen (22%) and 5 (8%) pts had a non-<i>BRCA</i> HRRm in the LT and ST groups, respectively, although 65% of pts with a non-<i>BRCA</i> HRRm had HRD-positive tumors. Twenty-five (37%) and 32 (49%) had <i>TP53</i>-disruptive mutations in the LT and ST groups, respectively. Overlap between t<i>BRCA</i>m, HRD, non-<i>BRCA</i> HRRm (including most frequently mutated genes), and <i>TP53</i> mutations is shown in the Figure. <b>Conclusions:</b> Pts with LT PFS (>18 mo) in OPINION more commonly had HRD-positive tumors, with or without a t<i>BRCA</i>m, than pts with ST PFS (<4 mo), consistent with results showing longer PFS among pts with HRD-positive tumors in the overall population. A higher proportion of pts with LT than ST PFS achieved CR to their last PBC. However, pts could achieve LT PFS regardless of clinical and molecular factors, reinforcing the benefit of maintenance PARP inhibition in non-g<i>BRCA</i>m PSR OC.

Single cell RNA-sequence analysis to identify transcriptomic differences associated with treatment outcome and ethnicity in circulating tumor cells (CTCs) from patients (pts) with metastatic colorectal cancer (mCRC).

3041 Background: CTCs are a promising diagnostic and prognostic biomarker in colorectal cancer. The analysis of CTCs at the single-cell level may allow to capture tumor heterogeneity and identify novel prognostic and predictive markers in mCRC. Our aim was to evaluate whether the transcriptome profile of CTCs was associated with progression-free survival (PFS) in pts receiving treatment for mCRC and whether we could identify significant differences based on pts’ ethnicity. Methods: Single CTCs were prospectively collected from 25 pts undergoing treatment for mCRC at the USC/Norris Comprehensive Cancer Center between April and October 2019. Pre-treatment peripheral blood was processed using CTC-FIND comprised filter separation and immunomagnetic depletion of CD45/50-expressing cells to collect ultra-pure CTCs samples. After isolation to single cell, CTCs were processed by single cell RNA-sequence (scRNAseq). Principal component analysis was used for clustering expression data. DESeq2 was used to identify differentially expressed genes between pts with short ( &lt; 150 days) and long term ( &gt; 150 days) PFS, as well as between Hispanic and non-Hispanic ethnicity with control for FDR ( Q &lt; 0.1). Ingenuity pathway analyses (IPA) of enriched pathway networks were performed. Results: Main pts characteristics were as follow: median age = 52 years; M/F = 18/7; Hispanics/non-Hispanics = 7/18; treatment line (n pts) = 1st (5), 2nd (11), ≥ 3rd (9). We identified 59 single CTC and CTC cluster from 22/25 pts (range 1-9, median = 2/pt). scRNAseq analysis identified two separate clusters of pts based on PFS (short term vs long term). Hispanic pts were mainly distributed within the short term PFS cluster. The IPA of the network of top 40 enriched pathways showed several pathways related with metabolism, such as Sirtuin signaling, mitochondrial dysfunction and oxidative phosphorylation, and IL6/JAK/STAT signaling pathways in pts with short term PFS ( Q &lt; 0.05). When comparing Hispanic vs non-Hispanic pts, we detected enrichment of neuroinflammation signaling pathway in Hispanics, including CXCL8 (fold change, FC: 11.04), GAD2 (FC: 4.77), IRAK3 (FC: 17.06), PLCG2 (FC: 4.65) and TYROBP (FC: 8.43) (all Q &lt; 0.05). Conclusions: In our study CTCs transcriptome profiles showed an association with PFS in pts receiving treatment for mCRC, with an enrichment in mitochondria-related pathways and IL6/JAK/STAT signaling in the CTCs of pts with shorter PFS. Additionally, CTCs scRNAseq identified differentially expressed genes in Hispanic pts, displaying enrichment in neuroinflammation signaling. These results highlight the potential of CTCs molecular profiling as a tool to identify novel prognostic and predictive biomarkers in mCRC and actionable molecular pathways which may impact tumor spread and treatment response

Preoperative chemotherapy compared with postoperative adjuvant chemotherapy for squamous cell carcinoma of the thoracic oesophagus with the detection of circulating tumour cells randomized controlled trial.

The role of preoperative chemotherapy in the treatment of patients with oesophageal squamous cell carcinoma (ESCC) remains controversial. Chemotherapy followed by surgery was compared with surgery±chemotherapy, and the detection of circulating tumour cells (CTCs) was performed on all enrolled patients. We randomly assigned patients with resectable tumours to the preoperative chemotherapy group (Pre group) or surgery group (patients who were either given or not given adjuvant chemotherapy according to their postoperative lymph node status, Post group). Blood samples were collected 1-3 days before treatment (including preoperative chemotherapy and surgery) and 7 days after surgery for CTC detection. From July 2016 to October 2018, 115 patients were enrolled in the study, of whom 57 were assigned to the Pre group and 58 to the Post group. The proportion of patients with stage III ESCC was 63.16% in the Pre group and 48.28% in the Post group. No patients died during chemotherapy. One patient exhibited a complete response to preoperative chemotherapy, and 13 patients exhibited partial responses. The 2-year progression-free survival (PFS) and overall survival (OS) rates were not significantly different between the Pre and Post groups. In the subgroup analysis, patients with CTC (+) prior to treatments receiving preoperative chemotherapy had a better 2-year PFS (71.90% vs. 38.73%, P=0.0379). In the Cox proportional hazards regression analysis, platelet count was proven to correlate significantly with disease progression (P=0.016), and no factors were proven to correlate significantly with mortality after the factors were balanced in the present analysis. Preoperative chemotherapy improved the short-term PFS when CTC detection was positive prior to any treatment for patients with stage II or III ESCC. CTC detection may be used as an index to guide individualized strategic decisions regarding preoperative chemotherapy, but more evidence is needed.

Abstract P1-02-03: The effect on short–term progression free survival of the detection of ≥4 FDG-avid nodes or occult N3–disease in breast cancer patients with PET/CT

Abstract Aim: The risk for locoregional recurrence (LRR), after neoadjuvant chemotherapy (NAC), is based on staging before NAC as well as the final pathology after NAC. Especially the number of tumour–positive axillary nodes, which is also an important selection factor for postoperative radiotherapy, is not adequately assessed by ultrasound before NAC or axillary lymph node dissection after NAC. PET/CT has a high positive predictive value for the detection of lymph node metastases. Newly found lymph node metastasis on PET/CT incentivized us to change the radiotherapy plan for patients with primary breast cancer scheduled for NAC in our institute. Koolen et al. reported an upstaging of 23% stage II–III breast cancer patients to the radiotherapy requiring high–risk group (≥4 FDG–avid axillary nodes or detection of occult N3–disease) due to new lymph node metastasis detected with FDG PET/CT imaging. In this study, we report the effect of this upstaging with PET/CT short–term progression free survival (PFS). Materials and methods: Between 2007 and 2011 a total of 278 breast cancer patients (mean age 48.9y, range 19–75y), with a tumour of at least 3 cm and without metastases, received a baseline PET/CT for staging purposes and subsequent response monitoring to NAC. The group was divided in three groups: a low– (T2N0), intermediate– (T0–2N1 and T3N0) and a high–risk group (T0–3N2–3, T3N1 and T4). We classified LRR, distant metastases and death as an "event"; including all patients in the PFS analysis of the first 3 years. Results: With a median follow–up (FU) of 37 months and the upstaging as depicted in table 1 Upstaging of breast cancer risk-group after PET/CTTotal group before PET/CTChanged After PET/CTComplete after PET/CT (FU-events)Low-risk: N=47N=5Low-risk (2) N= 42Intermediate-risk: N=144N=38Intermediate-risk N= 106 (14)High-risk: N=87xHigh-risk N= 130 (27)Total: N=278Total: N= 43Total N= 278Table 1: The group was divided in three groups: a low- (T2N0), intermediate- (T0-2N1 and T3N0) and a high-risk group (T0-3N2-3, T3N1 and T4). The table shows the upstaging of breast cancer patients from low- and intermediate- to the high-risk group, and thus, after PET/CT, requiring radiotherapy. : The patients not upstaged by PET/CT showed no difference in PFS between the high–risk, intermediate–risk and low–risk groups (Logrank p=0.18). Due to the migration of the 43 patients from the low– and intermediate group to the high–risk group, based on PET/CT findings, the PFS differed significantly between the risk–groups (Logrank p=0.04). No difference in loco–regional recurrence was seen between the low–risk and the high–risk group (P=0.18). Conclusion: After upstaging with PET/CT, into the high–risk group requiring radiotherapy, a significant difference is seen between the three risk–groups. PET/CT restaging may more adequately predict progression free survival. The detection occult lymphatic metastasis with PET/CT leads to upstaging in clinically unsuspected patients with primary breast cancer, enabling adequate radiotherapy treatment. Citation Format: Suzana C Teixeira, Bas B Koolen, Paula HM Elkhuizen, Vincent van der Noort, Marie-Jeanne Vrancken-Peeters, Marcel P Stokkel, Emiel J Th Rutgers, Renato A Valdés-Olmos. The effect on short–term progression free survival of the detection of ≥4 FDG-avid nodes or occult N3–disease in breast cancer patients with PET/CT [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P1-02-03.

18F-FDOPA and 18F-FLT positron emission tomography parametric response maps predict response in recurrent malignant gliomas treated with bevacizumab

The current study examined the use of voxel-wise changes in (18)F-FDOPA and (18)F-FLT PET uptake, referred to as parametric response maps (PRMs), to determine whether they were predictive of response to bevacizumab in patients with recurrent malignant gliomas. Twenty-four patients with recurrent malignant gliomas who underwent bevacizumab treatment were analyzed. Patients had MR and PET images acquired before and at 2 time points after bevacizumab treatment. PRMs were created by examining the percentage change in tracer uptake between time points in each image voxel. Voxel-wise increase in PET uptake in areas of pretreatment contrast enhancement defined by MRI stratified 3-month progression-free survival (PFS) and 6-month overall survival (OS) according to receiver-operating characteristic curve analysis. A decrease in PET tracer uptake was associated with longer PFS and OS, whereas an increase in PET uptake was associated with short PFS and OS. The volume fraction of increased (18)F-FDOPA PET uptake between the 2 posttreatment time points also stratified long- and short-term PFS and OS (log-rank, P < .05); however, (18)F-FLT uptake did not stratify OS. This study suggests that an increase in FDOPA or FLT PET uptake on PRMs after bevacizumab treatment may be a useful biomarker for predicting PFS and that FDOPA PET PRMs are also predictive of OS in recurrent gliomas treated with bevacizumab.

Inhibition of Usp9x Deubiquitinase Activity by WP1130 Reduces Mcl-1 Levels and Induces Apoptosis In Cells From Patients with Plasma Cell Dyscrasia and Drug-Refractory Multiple Myeloma

AbstractAbstract 3005Multiple myeloma (MM) patients are typically treated with a number of combinations of unconventional drugs that provide excellent and sometimes durable responses. However, most patients are expected to relapse or progress and relapsed disease is usually less sensitive to chemotherapy. Therefore, drug resistance remains a key concern in MM therapy. Some important clues regarding MM drug and apoptotic resistance have been obtained from biochemical and molecular studies of pro-survival genes. Mcl-1 is one member of the Bcl-2 family with strong anti-apoptotic activity. Mcl-1 binds pro-apoptotic proteins such as Bim and Bak, forming hetero-dimers to suppress apoptosis. Specific knockdown of Mcl-1 but not other anti-apoptotic Bcl-2 proteins (Bcl-2, Bcl-xL) induces apoptosis in MM cells and a critical level of Mcl-1 is essential to MM cell viability. Also, drug resistant MM patients are reported to express higher levels of Mcl-1 when compared to drug naïve patients. These observations suggest that Mcl-1 plays a key role in MM drug and apoptotic resistance. Mcl-1 is unique among survival proteins as its cellular protein level is primarily controlled by its ubiquitination and proteasomal degradation. Recently Usp9x, a high MW de-ubiquitinase (DUB), was shown to bind and stabilize Mcl-1 by mediating its de-ubiquitination and loss of recognition by the proteasome. Usp9x was also shown to be highly expressed in MM patients with short term progression-free survival. Analysis of Usp9x gene expression in MM patients treated at our center supports a correlation between elevated Usp9x expression (>4-fold over normal PBMCs) and poor patient prognosis. These observations suggest that agents that inhibit Usp9x activity may be effective in controlling Mcl-1 levels and have therapeutic impact in MM patients. Here we describe a small molecule Usp9x inhibitor, WP1130, that causes a rapid reduction in Mcl-1 protein levels and stimulates apoptosis in MM cells. We further demonstrate that the anti-tumor activity of WP1130 is mediated through inhibition of Usp9x and other active DUBs in B-cell tumors. WP1130-mediated inhibition of Usp9x activity promotes Mcl-1 ubiquitination leading to its rapid (1-4 hours) proteasomal degradation with subsequent induction of apoptosis. Pretreatment of MM cells with proteasome inhibitors (MG-132 or bortezomib) prior to WP1130 treatment blocked the down-regulation of Mcl-1, demonstrating proteasome involvement in regulation of Mcl-1 levels by WP1130. Primary MM tumor cells (bone marrow aspirates) derived from newly diagnosed and advanced therapy refractory patients were also screened for their responsiveness to WP1130 by DUB activity assays. Samples were obtained from a newly diagnosed plasmacytoma/plasma cell leukemia patient and 4 MM patients refractory to the Velcade/Doxorubicin/Dexamethasone (VDD) regimen. Usp9x activity was detected in 4 of 5 patient specimens and WP1130 treatment reduced Mcl-1 protein levels and induced apoptosis only in samples with active Usp9x. Furthermore, WP1130 completely eliminated CD138+ cells after 24 hours in MM patient specimens with active Usp9x while only an 11% reduction was measured in MM cells that did not exhibit Usp9x activity. Cytogenetic prognostic predictors (del13) did not distinguish between WP1130-responsive and non-responsive cells. Our results suggest that inhibition of Usp9x activity by WP1130 is associated with Mcl-1 down-regulation in MM cells. These results also suggest that drug-refractory MM patients with elevated Usp9x activity would likely benefit from Usp9x inhibitor-based therapy. Preliminary MM tumor studies have shown that WP1130 suppresses tumor growth in vivo and can be administered safely. Together, these results suggest that WP1130-mediated Usp9x inhibition may be an effective and novel approach in treating progressive or drug-refractory MM patients. Disclosures:Jakubowiak:Millenium, Celgene, Bristol-Myers Squibb, Johnson & Johnson Ortho-Centocor: Honoraria; Millennium, Bristol-Myers Squibb: Membership on an entity's Board of Directors or advisory committees; Millennium, Celgene, Centocor-Ortho Biotech: Speakers Bureau.

An ANOCEF genomic and transcriptomic microarray study of the response to radiotherapy or to alkylating first-line chemotherapy in glioblastoma patients.

BackgroundThe molecular characteristics associated with the response to treatment in glioblastomas (GBMs) remain largely unknown. We performed a retrospective study to assess the genomic characteristics associated with the response of GBMs to either first-line chemotherapy or radiation therapy. The gene expression (n = 56) and genomic profiles (n = 67) of responders and non-responders to first-line chemotherapy or radiation therapy alone were compared on Affymetrix Plus 2 gene expression arrays and BAC CGH arrays.ResultsAccording to Verhaak et al.'s classification system, mesenchymal GBMs were more likely to respond to radiotherapy than to first-line chemotherapy, whereas classical GBMs were more likely to respond to first-line chemotherapy than to radiotherapy. In patients treated with radiation therapy alone, the response was associated with differential expression of microenvironment-associated genes; the expression of hypoxia-related genes was associated with short-term progression-free survival (< 5 months), whereas the expression of immune genes was associated with prolonged progression-free survival (> 10 months). Consistently, infiltration of the tumor by both CD3 and CD68 cells was significantly more frequent in responders to radiotherapy than in non-responders. In patients treated with first-line chemotherapy, the expression of stem-cell genes was associated with resistance to chemotherapy, and there was a significant association between response to treatment and p16 locus deletions. Consistently, in an independent data set of patients treated with either radiotherapy alone or with both radiotherapy and adjuvant chemotherapy, we found that patients with the p16 deletion benefited from adjuvant chemotherapy regardless of their MGMT promoter methylation status, whereas in patients without the p16 deletion, this benefit was only observed in patients with a methylated MGMT promoter.ConclusionDifferential expression of microenvironment genes and p16 locus deletion are associated with responses to radiation therapy and to first-line chemotherapy, respectively, in GBM. Recently identified transcriptomic subgroups of GBMs seem to respond differently to radiotherapy and to first-line chemotherapy.

Analytical Approaches for the BOAC SNP Panel Association with Progression Free Survival in Myeloma

The Bank On A Cure (BOAC) has established DNA banks from multiple cooperative and institutional clinical trials, and platforms for examining the association of genetic variations (SNPs) with disease risk and outcomes in myeloma. We have previously described the development and content of a novel custom SNP panel that contains 3,404 SNPs in 983 genes, representing cellular functions and pathways that may influence disease response, toxicities, complications, and survival. Although survival certainly varies according to tumor heterogeneity (ie. chromosomal abnormalities, gene expression variations) germline variations that influence the microenvironment, drug distribution, drug transport and metabolism, may also have an association with event free survival outcomes. To explore SNP associations with progression free survival (PFS) we compared the BOAC SNP profiles of short term PFS (less than 1 year, n=70) versus long term PFS (greater than 3 years, n=73) in two phase III clinical trials (ECOG E9487 and SWOG S9321). A variety of analytical approaches was undertaken including univariate rank ordering, recursive partitioning, and support vector machine learning tools (SVM). Each of these approaches has advantages and limitations in dealing with type I false positive errors as well as sensitivity and specificity. We included subset validation approaches and randomization of classes to address how robust and predictive different approaches were. From our analysis we conclude germline genomic variations do have an impact on progression free survival, with a subset of SNPs from the panel reaching 76% predictive association and hazard ratios of PFS of 9.6 (CI 4.5, 20.5), p<0.001, using SVM analysis. Based on univariate approaches, we find the most significant variations associated with PFS differences were genes that could be functionally categorized as pharmacologic. The presentation will focus on the analytical approaches, and refinements necessary to assure predictive value compared to random associations. Notwithstanding the clear importance of tumor cell variations in genetic deregulation, we conclude that various functions within the bone marrow and drug response likely interplay as a complex influence on disease progression, response, and survival. This suggests combining gene expression profiles of the tumors with germline SNP profiles may provide more accurate prognosis. These combined analytical approaches are currently being developed with BOAC data bases, and examples will be discussed.