ALBERT

Description: Recently, several novel prognostic factors have been identified; their significance has been demonstrated in selected patient (pt) populations and retrospective analyses. As a group, previously treated pts with CLL likely have their respective, relevant prognostic factors for clinical endpoints, which may be further impacted by treatment (Rx). We prospectively evaluated the significance of newer prognostic factors: FISH abnormalities (abn) (Vysis CLL panel), IgVH mutation status, ZAP70 expression (flow & immunohistochemistry), CD38 expression (≥30%); as well as traditional factors: conventional cytogenetic analysis perfomed on bone marrow metaphases, age, sex, # prior Rx, refractoriness to alkylating agents (ALK) or fludarabine (FLU), absolute lymphocyte count (ALC), HGB, PLT, β-2 microglobulin (B2M), ALB, LDH, creatinine, and Alk Phos as independent predictors for survival in previously treated pts. The group included 473 previously treated pts seen at M.D.Anderson (10/03–8/07), who were evaluated by bone marrow sampling with conventional and FISH cytogenetic analyses, and the new and traditional prognostic factors described above. The median (range) age was 63yrs(31–87) and # prior Rx was 2(1–13). Other characteristics were: 43% Rai high-risk; 35% FLU-refractory; and 39% ALK-refractory; 74% unmutated IgVH; 54% ZAP70+ (flow); 76% ZAP70+ (IHC); and 68% CD38+. FISH results were: 22% del 17p13, 21% del 11q22, 10% +12, and 48% del 13q14 or no abn by the hierarchical classification. Conventional cytogenetic analysis of bone marrow metaphases demonstrated 25% with a complex karyotypic abn (〉1 cell with 〉1 chromosome abn), 58% diploid, 17% with single clonal abn (〉1 cell with 1 abn). Of the 100 pts with complex karyotypic abn, 50% had del 17p13, 28% del 11q22, 6% +12, 9% del 13q14, and 7% had no abn by FISH. Survival was measured from the time of prognostic factor characterization (FISH). The median follow-up time was 10mo(0–47). Univariate analyses identified the following significant (p≤.01) predictors for shorter survival: advanced age, # prior Rx, Rai high-risk, ALK- or FLU-refractory, FISH del 17p13; complex karyotypic abn (Figure 1), unmutated IgVH, high ALC, low HGB, low PLT, high B2M, low ALB, high LDH, and high Alk Phos. Multivariate analysis produced the following model with the following significant (p

Description: Background: Low oxygen levels are a defining characteristic of solid tumors, but the role of hypoxia in leukemogenesis remains unclear. Recent reports indicate that the endosteum at the murine bone-bone marrow (BM) interface is hypoxic, and data in a rat model demonstrate that leukemic cells infiltrating bone marrow were markedly hypoxic compared with cells in the BM of healthy rats. Hypoxia triggers a complex cellular and systemic adaptation mediated primarily through transcription by hypoxia inducible factors (HIFs) including HIF-1a. Although hypoxia is the best-characterized mechanism of HIF activation in tumors, HIF activity also can be induced in tumor cells through activation of the PI3K/Akt-signaling pathway. In this study, we assessed AKT and HIF-1a expression in newly diagnosed precursor B-cell acute lymphoblastic leukemia (pre-B ALL) and correlated the results with overall and progression-free patient survival. Methods: We analyzed expression of phosphorylated AKT (pAKT) and HIF-1a in leukemic cells by immunohistochemical methods using archival fixed, paraffin-embedded BM biopsy specimens of newly diagnosed pre-B ALL and antibodies specific for pAKT (Cell Signaling Technology, Beverly, MA) and HIF-1a (BD Biosciences, San Jose, CA). The initial observations were confirmed by a Reverse Phase Protein Array (RPPA) data set generated from protein lysates prepared from fresh blood and BM aspirate samples from patients with newly diagnosed pre-B ALL. Results: There were 26 men and 27 women with a median age of 39 years (range, 17–77). The median follow-up was 17 months (range, 1–71). The median WBC was 5.7 × 109/L (range, 0.8–369 × 109/L), the median percentage of blasts in bone marrow was 88% (range, 21–97%). Conventional cytogenetic studies detected a normal karyotype in 13 patients and abnormal karyotype in 37 patients including the Philadelphia chromosome (Ph) in 15 patients; no analyzable metaphases were recovered in 5 patients. Fluorescence in situ hybridization for BCR/ABL rearrangement was performed in all patients and was positive in all 15 patients with Ph and in 1 patient with normal conventional cytogenetics. 50 patients received HYPER-CVAD therapy, 3 patients received augmented BFM therapy. 49 (92%) patients achieved complete remission with a median time to response of 3 weeks (range, 2–8 weeks), 12 of them relapsed. 17 patients died, including 6 patients in complete remission. 3 year overall survival was 56% (CI, 46–66%). HIF-1a expression was detected in 37 (70%) patients, including 10 patients with Ph-positive ALL. HIF-1a expression was associated with expression of pAKT (R=0.4479, p

Description: In chronic myelogenous leukemia (CML), treatment of early stage disease has been significantly improved with the discovery of imatinib mesylate (IM, Gleevec), but with the progression of the disease, drug resistance develops which is due to mutation of the ATP binding site domain within the Bcr-Abl tyrosine kinase, the activation of Lyn kinase (a Src-related tyrosine kinase) and other mutations that produce a more aggressive disease. Thus, patients relapse and the disease progresses to blast crisis (BC) stage where survival is limited to only several months. No suitable drug treatment for BC is available, and the mechanism of drug resistance for BC is unclear. A new synthesized small molecule, WP1193, a caffeic acid derivative, induced apoptosis in all types of drug-resistant CML cells including IM-resistant T315I BCR-ABL+ cells, IM-resistant CML cells with up-regulated Lyn tyrosine kinase (K562-R) and cells from blast crisis CML patients. Treatment of BCR-ABL+ cells with WP1193 inhibited Janus kinase activities, leading to strong reduction of Bcr-Abl; this treatment also reduced STAT3 and pTyr 705 STAT3 protein levels. CML cell lines contain a large signaling network complex composed of Bcr-Abl, Jak2, HSP90 and its client proteins. HSP90 is a molecular chaperone that plays an important role in conformational maturation and stabilization of client proteins (Bcr- Abl, Jak2, STAT3, etc.), which are physically associated in the signaling network complex. This network complex (estimated to be between 2–5 million daltons) has been characterized by gel filtration column chromatography. This complex is disassembled by treatment of Bcr-Abl+ cells with WP1193 (10 μM for 3–6 h). Treatment of the cells with WP1193 also inhibited binding of STAT3 to its consensus DNA sequence and reduced levels of HSP90α transcripts, HSP90α protein and its client proteins, which include Bcr-Abl, Jak2 and Akt. WP1193 also reduced solid tumors in mice inoculated with IM-resistant K562-R cells, and inhibited leukemic effects caused by T315I BCR-ABL+ cells. These results indicate that disruption of the Bcr-Abl/Jak2/HSP90 network complex by WP1193 overcomes drug resistance in advanced stage CML cells. Since disruption of this network complex causes apoptosis induction in all IM-resistant and late stage CML patient cells tested so far, and since no significant toxicity was observed in short-term mouse experiments, WP1193 may have medical utility for the treatment of IM-resistant forms of CML and late stage CML where other drugs fail. One novel feature of WP1193 is its ability to reduce the activity of several important components of the network, namely Bcr-Abl, Jak2 and STAT3 activities and reduction of HSP90α, which then causes a high level of apoptosis in IM-resistant cells and late-stage CML cells.