Unraveling the complexity of tyrosine kinase inhibitor-resistant populations by ultra-deep sequencing of the BCR-ABL kinase domain

In chronic myeloid leukemia and Philadelphia chromosome-positive acute lymphoblastic leukemia, tyrosine kinase inhibitor (TKI) therapy may select for drug-resistant BCR-ABL mutants. We used an ultra-deep sequencing (UDS) approach to resolve qualitatively and quantitatively the complexity of mutated populations surviving TKIs and to investigate their clonal structure and evolution over time in relation to therapeutic intervention. To this purpose, we performed a longitudinal analysis of 106 samples from 33 patients who had received sequential treatment with multiple TKIs and had experienced sequential relapses accompanied by selection of 1 or more TKI-resistant mutations. We found that conventional Sanger sequencing had misclassified or underestimated BCR-ABL mutation status in 55% of the samples, where mutations with 1% to 15% abundance were detected. A complex clonal texture was uncovered by clonal analysis of samples harboring multiple mutations and up to 13 different mutated populations were identified. The landscape of these mutated populations was found to be highly dynamic. The high degree of complexity uncovered by UDS indicates that conventional Sanger sequencing might be an inadequate tool to assess BCR-ABL kinase domain mutation status, which currently represents an important component of the therapeutic decision algorithms. Further evaluation of the clinical usefulness of UDS-based approaches is warranted

Implant rehabilitation of the edentulous jaws: Does tilting of posterior implants at an angle greater than 45\ub0 affect bone resorption and implant success?: A retrospective study

Purpose: This study aimed to (1) investigate the success of posterior implants tilted >45\ub0 when 4 immediately loaded implants were used to support full-arch prostheses, eliminating any distal cantilever and (2) examine the effect on marginal bone loss (MBL) of different combinations of anterior multi-unit abutment (MUA) angles and posterior implant tilting angles. Materials and Methods: Records of patients rehabilitated according to the Columbus Bridge Protocol were analyzed. Peri-implant bone levels (PBLs) and MBL were measured for each implant. The influence of posterior implant tilting angle on PBL, MBL, and implant and prosthetic success rate was investigated. The impact on the same endpoints of different anterior MUA angles, and different combinations of anterior MUA and tilted posterior implant angles was also examined. Results: Records of 41 patients were analyzed, for a total of 46 complete rehabilitations, and 142 implants (52 anterior, 63 posterior tilted 6445\ub0 [group 1], and 27 posterior tilted >45\ub0 [group 2]). No implants were lost during the follow-up (25.9 months), and no prosthetic complications were reported. Success rate for posterior implants was 100% in group 1 and 96.3% in group 2. Mean MBL differed significantly between the 2 groups (0.45 mm in group 1, 0.66 in group 2 [P =.04]), but not when the analysis was limited to implants in the same jaw. Implant tilting angle did not correlate with MBL and the MUA angle had no effect on bone resorption around posterior implants, neither in the sample as a whole nor in individual patients. Conclusions: Posterior implants tilted >45\ub0 to eliminate distal cantilever may be as safe as those tilted less in severely atrophic jaws rehabilitated with immediately loaded, full-arch prostheses supported on 4 implants. Further prospective studies on larger samples of patients and implants and with longer follow-up are needed to confirm these findings

Prospective phase II Study on 5-days azacitidine for treatment of symptomatic and/or erythropoietin unresponsive patients with low/INT-1-risk myelodysplastic syndromes.

Purpose: This phase II prospective study aimed to evaluate the efficacy and safety of 5-days azacytidine (5d-AZA) in patients with low-risk myelodysplastic syndromes (MDS). Second, single-nucleotide polymorphism (SNP) genetic profile and phosphoinositide-phospholipase C (PI-PLC) β1 levels were studied to evaluate possible biologic markers able to predict the hematologic response. Experimental Design: The study tested a lower intensity schedule of azacytidine. The treatment plan consisted of 75 mg/sqm/d subcutaneous administered for 5 days every 28 days, for a total of 8 cycles. Results: Thirty-two patients were enrolled in the study. The overall response rate was 47% (15 of 32) on intention-to-treat and 58% (15 of 26) for patients completing the treatment program. In this latter group, 5 (19%) achieved complete remission (CR) and 10 (38%) had hematologic improvement, according to the International Working Group (IWG) criteria. Three patients have maintained their hematologic improvement after 37, 34, and 33 months without other treatments. Moreover, 21 and 2 of 26 cases completing 8 cycles were transfusion-dependent for red blood cells and platelets at baseline, respectively. Of these, 7 (33%) and 2 (100%) became transfusion-independent at the end of the treatment program, respectively. Grade 3-4 neutropenia occurred in 28% of patients and 4 patients died early due to infections or hemorrhage. SNP results were not significantly correlated to the clinical outcome, whereas PI-PLCβ1 level anticipated either positive or negative clinical responses. Conclusions: 5d-AZA is safe and effective in a proportion of patients with low-risk MDS. PI-PLCβ1 gene expression is a reliable and dynamic marker of response that can be useful to optimize azacytidine therapy. © 2013 American Association for Cancer Research