Jonathan L. Jesneck

H3K79 methylation profiles define murine and human MLL-AF4 leukemias

We created a mouse model wherein conditional expression of an Mll-AF4 fusion oncogene induces B precursor acute lymphoblastic (ALL) or acute myeloid leukemias (AML). Gene expression profile analysis of the ALL cells demonstrated significant overlap with human MLL-rearranged ALL. ChIP-chip analysis demonstrated histone H3 lysine 79 (H3K79) methylation profiles that correlated with Mll-AF4-associated gene expression profiles in murine ALLs and in human MLL-rearranged leukemias. Human MLL-rearranged ALLs could be distinguished from other ALLs by their H3K79 profiles, and suppression of the H3K79 methyltransferase DOT1L inhibited expression of critical MLL-AF4 target genes. We thus demonstrate that ectopic H3K79 methylation is a distinguishing feature of murine and human MLL-AF4 ALLs and is important for maintenance of MLL-AF4-driven gene expression.

The OTT-MAL fusion oncogene activates RBPJ-mediated transcription and induces acute megakaryoblastic leukemia in a knockin mouse model

Acute megakaryoblastic leukemia (AMKL) is a form of acute myeloid leukemia (AML) associated with a poor prognosis. The genetics and pathophysiology of AMKL are not well understood. We generated a knockin mouse model of the one twenty-two-megakaryocytic acute leukemia (OTT-MAL) fusion oncogene that results from the t(1;22)(p13;q13) translocation specifically associated with a subtype of pediatric AMKL. We report here that OTT-MAL expression deregulated transcriptional activity of the canonical Notch signaling pathway transcription factor recombination signal binding protein for immunoglobulin kappa J region (RBPJ) and caused abnormal fetal megakaryopoiesis. Furthermore, cooperation between OTT-MAL and an activating mutation of the thrombopoietin receptor myeloproliferative leukemia virus oncogene (MPL) efficiently induced a short-latency AMKL that recapitulated all the features of human AMKL, including megakaryoblast hyperproliferation and maturation block, thrombocytopenia, organomegaly, and extensive fibrosis. Our results establish that concomitant activation of RBPJ (Notch signaling) and MPL (cytokine signaling) transforms cells of the megakaryocytic lineage and suggest that specific targeting of these pathways could be of therapeutic value for human AMKL.

The Apcmin mouse has altered hematopoietic stem cell function and provides a model for MPD/MDS

Apc, a negative regulator of the canonical Wnt signaling pathway, is a bona-fide tumor suppressor whose loss of function results in intestinal polyposis. APC is located in a commonly deleted region on human chromosome 5q, associated with myelodysplastic syndrome (MDS), suggesting that haploinsufficiency of APC contributes to the MDS phenotype. Analysis of the hematopoietic system of mice with the Apc(min) allele that results in a premature stop codon and loss of function showed no abnormality in steady state hematopoiesis. Bone marrow derived from Apc(min) mice showed enhanced repopulation potential, indicating a cell intrinsic gain of function in the long-term hematopoietic stem cell (HSC) population. However, Apc(min) bone marrow was unable to repopulate secondary recipients because of loss of the quiescent HSC population. Apc(min) mice developed a MDS/myeloproliferative phenotype. Our data indicate that Wnt activation through haploinsufficiency of Apc causes insidious loss of HSC function that is only evident in serial transplantation strategies. These data provide a cautionary note for HSC-expansion strategies through Wnt pathway activation, provide evidence that cell extrinsic factors can contribute to the development of myeloid disease, and indicate that loss of function of APC may contribute to the phenotype observed in patients with MDS and del(5q).

Antigen Availability Determines CD8+ T Cell-Dendritic Cell Interaction Kinetics and Memory Fate Decisions

T cells are activated by antigen (Ag)-bearing dendritic cells (DCs) in lymph nodes in three phases. The duration of the initial phase of transient, serial DC-T cell interactions is inversely correlated with Ag dose. The second phase, characterized by stable DC-T cell contacts, is believed to be necessary for full-fledged T cell activation. Here we have shown that this is not the case. CD8⁺ T cells interacting with DCs presenting low-dose, short-lived Ag did not transition to phase 2, whereas higher Ag dose yielded phase 2 transition. Both antigenic constellations promoted T cell proliferation and effector differentiation but yielded different transcriptome signatures at 12 hr and 24 hr. T cells that experienced phase 2 developed long-lived memory, whereas conditions without stable contacts yielded immunological amnesia. Thus, T cells make fate decisions within hours after Ag exposure, resulting in long-term memory or abortive effector responses, correlating with T cell-DCs interaction kinetics.

Physical characterization of a prototype selenium-based full field digital mammography detector.

The purpose of this study was to measure experimentally the physical performance of a prototype mammographic imager based on a direct detection, flat-panel array design employing an amorphous selenium converter with 70 microm pixels. The system was characterized for two different anode types, a molybdenum target with molybdenum filtration (Mo/Mo) and a tungsten target with rhodium filtration (W/Rh), at two different energies, 28 and 35 kVp, with approximately 2 mm added aluminum filtration. To measure the resolution, the presampled modulation transfer function (MTF) was measured using an edge method. The normalized noise power spectrum (NNPS) was measured by two-dimensional Fourier analysis of uniformly exposed mammograms. The detective quantum efficiencies (DQEs) were computed from the MTFs, the NNPSs, and theoretical ideal signal to noise ratios. The MTF was found to be close to its ideal limit and reached 0.2 at 11.8 mm(-1) and 0.1 at 14.1 mm(-1) for images acquired at an RQA-M2 technique (Mo/Mo anode, 28 kVp, 2 mm Al). Using a tungsten technique (MW2; W/Rh anode, 28 kVp, 2 mm Al), the MTF went to 0.2 at 11.2 mm(-1) and to 0.1 at 13.3 mm(-1). The DQE reached a maximum value of 54% at 1.35 mm(-1) for the RQA-M2 technique at 1.6 microC/kg and achieved a peak value of 64% at 1.75 mm(-1) for the tungsten technique (MW2) at 1.9 microC/kg. Nevertheless, the DQE showed strong exposure and frequency dependencies. The results indicated that the detector offered high MTFs and DQEs, but structured noise effects may require improved calibration before clinical implementation.

Fundamental imaging characteristics of a slot-scan digital chest radiographic system

Our purpose in this study was to evaluate the fundamental image quality characteristics of a new slot-scan digital chest radiography system (ThoraScan, Delft Imaging Systems/Nucletron, Veenendaal, The Netherlands). The linearity of the system was measured over a wide exposure range at 90, 117, and 140 kVp with added Al filtration. System uniformity and reproducibility were established with an analysis of images from repeated exposures. The modulation transfer function (MTF) was evaluated using an established edge method. The noise power spectrum (NPS) and the detective quantum efficiency (DQE) of the system were evaluated at the three kilo-voltages over a range of exposures. Scatter fraction (SF) measurements were made using a posterior beam stop method and a geometrical chest phantom. The system demonstrated excellent linearity, but some structured nonuniformities. The 0.1 MTF values occurred between 3.3-3.5 mm(-1). The DQE(0.15) and DQE(2.5) were 0.21 and 0.07 at 90 kVp, 0.18 and 0.05 at 117 kVp, and 0.16 and 0.03 at 140 kVp, respectively. The system exhibited remarkably lower SFs compared to conventional full-field systems with anti-scatter grid, measuring 0.13 in the lungs and 0.43 in the mediastinum. The findings indicated that the slot-scan design provides marked scatter reduction leading to high effective DQE (DQEeff) of the system and reduced patient dose required to achieve high image quality.

TCR-dependent transformation of mature memory phenotype T cells in mice

A fundamental goal in cancer research is the identification of the cell types and signaling pathways capable of initiating and sustaining tumor growth, as this has the potential to reveal therapeutic targets. Stem and progenitor cells have been implicated in the genesis of select lymphoid malignancies. However, the identity of the cells in which mature lymphoid neoplasms are initiated remains unclear. Here, we investigate the origin of peripheral T cell lymphomas using mice in which Snf5, a chromatin remodelling-complex subunit with tumor suppressor activity, could be conditionally inactivated in developing T cells. In this model of mature peripheral T cell lymphomas, the cell of origin was a mature CD44hiCD122loCD8⁺ T cell that resembled a subset of memory cells that has capacity for self-renewal and robust expansion, features shared with stem cells. Further analysis showed that Snf5 loss led to activation of a Myc-driven signaling network and stem cell transcriptional program. Finally, lymphomagenesis and lymphoma proliferation depended upon TCR signaling, establishing what we believe to be a new paradigm for lymphoid malignancy growth. These findings suggest that the self-renewal and robust proliferative capacities of memory T cells are associated with vulnerability to oncogenic transformation. Our findings further suggest that agents that impinge upon TCR signaling may represent an effective therapeutic modality for this class of lethal human cancers.

Do Serum Biomarkers Really Measure Breast Cancer

BackgroundBecause screening mammography for breast cancer is less effective for premenopausal women, we investigated the feasibility of a diagnostic blood test using serum proteins.MethodsThis study used a set of 98 serum proteins and chose diagnostically relevant subsets via various feature-selection techniques. Because of significant noise in the data set, we applied iterated Bayesian model averaging to account for model selection uncertainty and to improve generalization performance. We assessed generalization performance using leave-one-out cross-validation (LOOCV) and receiver operating characteristic (ROC) curve analysis.ResultsThe classifiers were able to distinguish normal tissue from breast cancer with a classification performance of AUC = 0.82 ± 0.04 with the proteins MIF, MMP-9, and MPO. The classifiers distinguished normal tissue from benign lesions similarly at AUC = 0.80 ± 0.05. However, the serum proteins of benign and malignant lesions were indistinguishable (AUC = 0.55 ± 0.06). The classification tasks of normal vs. cancer and normal vs. benign selected the same top feature: MIF, which suggests that the biomarkers indicated inflammatory response rather than cancer.ConclusionOverall, the selected serum proteins showed moderate ability for detecting lesions. However, they are probably more indicative of secondary effects such as inflammation rather than specific for malignancy.

Computer-aided classification of breast microcalcification clusters: merging of features from image processing and radiologists

We developed an ensemble classifier for the task of computer-aided diagnosis of breast microcalcification clusters,which are very challenging to characterize for radiologists and computer models alike. The purpose of this study is to help radiologists identify whether suspicious calcification clusters are benign vs. malignant, such that they may potentially recommend fewer unnecessary biopsies for actually benign lesions. The data consists of mammographic features extracted by automated image processing algorithms as well as manually interpreted by radiologists according to a standardized lexicon. We used 292 cases from a publicly available mammography database. From each cases, we extracted 22 image processing features pertaining to lesion morphology, 5 radiologist features also pertaining to morphology, and the patient age. Linear discriminant analysis (LDA) models were designed using each of the three data types. Each local model performed poorly; the best was one based upon image processing features which yielded ROC area index AZ of 0.59 ± 0.03 and partial AZ above 90% sensitivity of 0.08 ± 0.03. We then developed ensemble models using different combinations of those data types, and these models all improved performance compared to the local models. The final ensemble model was based upon 5 features selected by stepwise LDA from all 28 available features. This ensemble performed with AZ of 0.69 ± 0.03 and partial AZ of 0.21 ± 0.04, which was statistically significantly better than the model based on the image processing features alone (p<0.001 and p=0.01 for full and partial AZ respectively). This demonstrated the value of the radiologist-extracted features as a source of information for this task. It also suggested there is potential for improved performance using this ensemble classifier approach to combine different sources of currently available data.

Ott1 (Rbm15) Is Essential for Placental Vascular Branching Morphogenesis and Embryonic Development of the Heart and Spleen

ABSTRACT The infant leukemia-associated gene Ott1 (Rbm15) has broad regulatory effects within murine hematopoiesis. However, germ line Ott1 deletion results in fetal demise prior to embryonic day 10.5, indicating additional developmental requirements for Ott1. The spen gene family, to which Ott1 belongs, has a transcriptional activation/repression domain and RNA recognition motifs and has a significant role in the development of the head and thorax in Drosophila melanogaster. Early Ott1-deficient embryos show growth retardation and incomplete closure of the notochord. Further analysis demonstrated placental defects in the spongiotrophoblast and syncytiotrophoblast layers, resulting in an arrest of vascular branching morphogenesis. The rescue of the placental defect using a conditional allele with a trophoblast-sparing cre transgene allowed embryos to form a normal placenta and survive gestation. This outcome showed that the process of vascular branching morphogenesis in Ott1-deficient animals was regulated by the trophoblast compartment rather than the fetal vasculature. Mice surviving to term manifested hyposplenia and abnormal cardiac development. Analysis of global gene expression of Ott1-deficient embryonic hearts showed an enrichment of hypoxia-related genes and a significant alteration of several candidate genes critical for cardiac development. Thus, Ott1-dependent pathways, in addition to being implicated in leukemogenesis, may also be important for the pathogenesis of placental insufficiency and cardiac malformations.