Clinical trials of monoclonal antibodies in combination with chemotherapy have reported previously unattained response rates in patients with B-cell chronic lymphocytic leukemia (B-CLL); however, the analysis of ZAP-70 protein and/or CD38 may explain better the discordant outcomes independent of treatment

Nucleophosmin-1 (NPM1) mutations represent the most frequent gene alteration in acute myeloid leukemia (AML). The most common NPM1 mutation type, accounting for 75 to 80% of cases , is referred to as mutation A (NPM1-mutA). These NPM1 alterations have been shown to possess prognostic significance because they appear to identify patients who will benefit from chemotherapy. Given the high prevalence and stability of these mutations over the course of disease , NPM1 mutations may serve as ideal targets for minimal residual disease (MRD) assessment in AML. Current detection methods are costly , require sophisticated equipment , and are often not sufficiently sensitive. We report here an allele-specific (ASO)-RT-PCR assay that enables rapid and sensitive detection of NPM1-mutA. A semi-nested ASO-PCR method was also designed to increase the sensitivity of our assay for the monitoring of MRD. We analyzed bone marrow cells collected from 52 patients with AML at presentation. NPM1-mutA was detected in leukemic cells from 21 patients. Nucleophosmin-1 (NPM1) is a nucleocytoplasmic shuttling protein that plays a key role in a variety of cellular processes including promotion of ribosome biogenesis, maintenance of genomic stability, regulation of transcription, and modulation of tumor-suppressor transcription factors.1 The NPM1 gene is one of the most frequent targets of genetic alterations in hematopoietic tumors as it has been shown to be involved in several gene fusions resulting from chromosome translocations and in other subtle mutations detected in lymphomas and acute leukemia. 2In early 2005, Falini et al 3 reported that NPM1 gene mutations, mainly consisting of small nucleotide insertion/ deletions, occur frequently in acute myeloid leukemia (AML) and are strongly associated with normal karyotype. Over 40 different types of mutations in the NPM1 locus have been described so far, which result in the formation of different mutant proteins. These alterations consist of the insertion/deletion of short nucleotide stretches (4 or 10 bp) at positions 956 through 971 in NPM1 exon 12 that lead to an open reading frame shift resulting in different protein variants containing novel C terminus portions.3-5 The most common NPM1 mutation type, accounting for 75% to 80% of cases, is referred to as mutation A (NPM1-mutA) 3 and consists of a duplication of a TCTG tetranucleotide at position 956 to 959 of the reference sequence (GenBank accession number NM_002520).

Myeloid sarcoma (MS, previously named granulocytic sarcoma or chloroma) is a rare extramedullary tumour of immature myeloid cells. It can be present before, concurrently with, or after the diagnosis of acute myeloid leukemia. MS is extremely uncommon in acute promyelocytic leukemia (APL). In the case described here, MS was the sole site of APL relapse and the cause of spinal cord compression. The patient presented with neurologic symptoms due to a paravertebral mass of MS after 7 years of complete remission. He was treated with excision of the mass followed by local radiotherapy. Systemic treatment was also given with combined arsenic trioxide and all-trans retinoic acid and the patient was able to achieve a second prolonged clinical and molecular remission.

Background: Trastuzumab and pertuzumab have been shown to work by inhibiting the intracellular signaling linked to HER2 receptor activation, and cytotoxic immune mechanisms, including Fc-dependent immune cell activation. ER+ and ER−/HER2+ breast cancers (BC) are considered molecularly distinct entities (Bianchini G, ASCO 2011; Iwamoto T, ESMO 2012). Regardless of the ER status, immune gene signatures are prognostic and predictive of chemotherapy response (Iwamoto T, ESMO 2012). The gene expression profiles of pre-treatment tumor samples from patients of the NeoSphere trial of neoadjuvant pertuzumab+trastuzumab ± docetaxel (T) (Gianni L, Lancet Oncol 2012) were characterized. We assessed the association of pre-selected adaptive immune functions and key immune regulatory genes with the likelihood of achieving a pathological complete response (pCR) in NeoSphere. Methods: Baseline core biopsies were collected from 387/417 patients. Gene expression profiles (Affymetrix U133 Plus 2.0) were obtained for 367 samples (88% of all patients) that were evenly distributed among arms A (TH, n=90), B (THP, n=95), C (HP, n=98), and D (TP, n=84). ER status was defined by IHC. The primary endpoint was pCR in breast. We assessed metagenes (average expression of the associated genes) corresponding to: immunoglobulins (IgG), CD8A, MHC type I and type II (MHC1, MHC2), interferon inducible genes (I-IG) and STAT1-related genes. We also assessed the individual genes PD1, PD-L1, PD-L2, CTLA4, IFNG. Results: ER+ and ER− tumors had differential mRNA expression for: CD8A, IgG, PD-L2, PD1, IFNG (overexpressed in ER−), and I-IG (overexpressed in ER+). Positive correlations were observed between most of these biomarkers. Only PD1 was inversely correlated with STAT1, interferon and MHC2. In logistic univariate regression analyses some biomarkers showed moderate association with pCR or residual disease without consistent patterns between treatment arms. However, a multivariate logistic regression model constructed for all the selected biomarkers revealed that high expression of PD-L1 was consistently associated with lower pCR rate in all chemotherapy containing arms (A, B and D). A similar trend was present also in Arm C, the arm with antibody treatment alone. In all arms, high expression of IFNG and/or STAT1 were associated with higher pCR rate. In multivariate analysis PD-L1, PD1 and STAT1 were associated with pCR irrespective of the ER status in combined arms A, C and D. In arm B, both CTLA4 and PD-L1 were independently associated with lower pCR in ER− tumors. Conclusions: The association between pCR and selected immune biomarkers in patients treated with pertuzumab, trastuzumab, or both, with or without chemotherapy, supports our understanding of the key role of the immune system in contributing to HER2-targeted antibody therapy on top of signaling inhibition. High PD-L1 expression emerged for its consistent association with lower pCR. These results provide a rationale for combining HER2-targeted treatments with immune-modulating agents and may allow the prediction of treatment benefit. Validation of these results with different assays is ongoing. Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr S6-7.

Summary. Concurrent resistance mechanisms, such as P-glycoprotein (PGP) and bcl-2, may contribute to a worse outcome in adult acute lymphoblastic leukaemia (ALL). Between 1990 and 2000, we analysed PGP and bcl-2 by flow cytometry, using two anti-PGP (C219 and JSB-1) monoclonal antibodies (mAbs) and an anti-bcl-2 mAb in 115 de novo adult ALL patients. Both a longer overall survival (OS) and longer disease-free survival (DFS) were observed in PGP-negative patients (23% vs 0% at 3 years, P ¼ 0AE011 and 29% vs 0% at 2 years, P ¼ 0AE006 for C219 respectively; 42% vs 0% at 1AE5 years, P ¼ 0AE004 and 53% vs 0% at 8AE5 months, P ¼ 0AE00006 for JSB-1 respectively). Bcl-2 positivity was associated with a significantly higher complete remission rate (90% vs 66%, P ¼ 0AE01). Moreover, in 69 patients not presenting with either t(9;22) or B-mature immunophenotype, PGP negativity (JSB-1) maintained its significant favourable prognostic impact with regard to OS (41% vs 0% at 1AE5 years, P ¼ 0AE009) and DFS (83% vs 0% at 6 months, P ¼ 0AE0005). Importantly, within a subset of 62 patients with normal (n ¼ 31) or unknown (n ¼ 31) karyotype, PGP (JSB-1)-negative patients showed both a significantly longer OS and DFS (63% vs 0% at 1AE4 years, P ¼ 0AE018 and 84% vs 0% at 6 months, P ¼ 0AE001 respectively). In multivariate analysis, JSB-1 (P ¼ 0AE008) and cytogenetics (P ¼ 0AE02) were found to be independent prognostic factors with regard to DFS. Therefore, in adult ALL, PGP and bcl-2 represent sensitive indicators of clinical outcome, and potential targets of novel molecules aimed at overcoming chemoresistance and recurrent relapses.