The purpose of this study was to determine the effects of interferon -(IFN -) gene transfer on the growth of PC3MM2 human prostate cancer cells in nude mice. Intralesional delivery of an adenoviral vector encoding murine IFN -( AdIFN -) , but not a vector encoding bacterial -galactosidase ( AdLacZ ) , suppressed PC3MM2 tumors in a dose -dependent manner. At the highest dose ( 2Â10 9 plaque -forming units, PFU ) , a single injection of AdIFN -( but not AdLacZ ) suppressed orthotopic PC3MM2 tumors and development of metastasis by 80%, and eradicated the tumors in 20% of mice. Immunohistochemical staining showed that AdIFN -± treated tumors contained fewer microvessels, fewer proliferating cells, and more apoptotic cells than did the control tumors. Compared with controls, tumors injected with AdIFN -expressed higher levels of IFN -and inducible nitric oxide synthase ( iNOS ) and lower levels of basic fibroblast growth factor ( bFGF ) and transforming growth factor 1 ( TGF -1). In vitro analysis indicated that expression of bFGF and TGF -1 in PC3MM2 cells could be suppressed by the nitric oxide donor sodium nitroprusside. These data suggest that intratumoral delivery of the IFN -gene with adenoviral vectors could be an effective therapy for prostate cancer and that tumor suppression by AdIFN -correlated with up -regulation of iNOS and down -regulation of angiogenesis.

Dysregulation of the phosphatidylinositol-3-kinase (PI3K) pathway in a wide range of tumors has made PI3K a consensus target to inhibit as illustrated by more than 15 inhibitors now in clinical trials. Our previous work, built on the early pioneering multikinase inhibitor LY294002, resulted in the only PI3K vascular-targeted PI3K inhibitor prodrug, SF1126, which has now completed Phase I clinical trials. This inhibitor has properties that impart more in vivo activity than should be warranted by its enzymatic potency, which in general is much lower than other clinical stage PI3K inhibitors. We embarked on the exploration of scaffolds that retained such properties while simultaneously exhibiting an increased potency toward PI3K. This work resulted in the discovery of the 5-morpholino-7H-thieno[3,2-b]pyran-7-one system as the foundation of a new compound class of potential PI3K inhibitors having improved potency toward PI3K. The synthesis and cancer stem cell-based activity of these compounds are reported herein.

Numerical change of chromosomes is common in acute myeloid leukemia (AML). However, a chromosome number as high as near-tetraploidy is very rare, especially in minimally differentiated AML (AML-M0). Erythrophagocytosis by reactive or malignant histiocytes is common in malignant hematological diseases; however, erythrophagocytosis by leukemic blasts is also very rare, especially in AML-M0. We report here the first case of AML-M0 with both of these unique characteristics: a near-tetraploid karyotype and erythrophagocytosis by leukemic blasts.

Introduction: Acute myeloid leukemia (AML) developing in chronic lymphocytic leukemia (CLL) patients is very uncommon and is usually treatment-related. Acute promyelocytic leukemia (APL) occurring concurrently with CLL is extremely rare and there is only one published case of treatment-related APL. No case of concomitant APL and CLL in patients without history of malignancy has been found in the literature. We report such a case of coexisting CLL and APL. Case report: A 52-years-old Caucasian male with past medical history of hypertension, diabetes mellitus, and coronary artery disease, presented to an outside facility with chest pain and dyspnea for two day duration. He had lymphocytosis (WBC 31,500 /µL and 66 % lymphocytes) one year prior, however, was lost to follow up and no further workup was pursued. On examination, he was pale with no petechiae or ecchymosis. There was no lymphadenopathy or hepatosplenomegaly. Labs showed WBC 17,000/µL with 97% lymphocytes and 2% blasts, Hemoglobin 6.0 g/dL, and platelet 16,000/µL. Serum troponin was 0.30 ng/mL. EKG revealed ST-depression in lateral leads concerning for non-ST elevation MI (NSTEMI). Patient was admitted and received packed red blood cells and platelet transfusion with resolution of chest pain. Upon transfer to our hospital, he was afebrile and hemodynamically stable. Lab work showed WBC of 72,000/µL and platelet count of 9,000/µL. Slides from outside facility were reviewed and immunohistochemistry from bone marrow specimen revealed dense staining of myeloperoxidase (MPO) on sheets of myeloid blasts and strong CD79a staining on clumps of lymphocytes. Peripheral blood, bone marrow aspiration and biopsy showed two distinct morphological abnormalities including immature myeloid blasts with prominent cytoplasmic granules as well as increased number of small lymphocytes. Flow cytometry demonstrated a clonal B-cell population consistent with chronic lymphocytic leukemia. Further evaluation with fluorescent in-situ hybridization (FISH) revealed presence of a t(15;17) (q22;q12), PML-RARA translocation consistent with acute promyelocytic leukemia. Co-existence of CLL and APL was confirmed. Treatment with All-trans-retinoic acid (ATRA) and hydroxyurea was started immediately along with dexamethasone for prevention of differentiation syndrome. Shortly after initiation of ATRA, patient developed acute hypoxic respiratory failure with extensive patchy opacities in bilateral lungs on chest radiography. WBC further increased to 130,000/µL. Respiratory failure worsened despite bi-level positive airway pressure (BiPAP) and diuretics, and he was subsequently transferred to intensive care unit (ICU). Patient's clinical condition deteriorated rapidly, developed disseminated intravascular coagulation (DIC), and eventually died from cardiopulmonary arrest. Discussion: CLL is the most common hematologic malignancy in adults in western countries and the treatment of CLL is associated with increased incidence of secondary malignancies. However, transformation of CLL into AML is uncommon and most reported cases were therapy-related (t-AML). In patients with t-AML, exposure to topoisomerase II inhibitors (mainly Etoposide), alkylating agents and ionizing radiation are among the main causative factors. AML after treatment with DNA-topoisomerase II inhibitors has a short latency period, presents without a prior myelodysplastic syndrome, and is associated with 11q23 translocation. There was only one reported case of APL which developed 2 years after radiotherapy for prostate cancer in a patient with chronic lymphocytic leukemia. Although no treatment was given for CLL, radiotherapy for prostate cancer in that patient might have contributed to the development of APL, which is considered therapy-related. High dose of radiation has also been considered to increase the risk of t-AML. To date, no report in literature has been found on simultaneous occurrence of CLL and APL in patients without any previous treatment, either for CLL or for other co-existing conditions. We report here the first case of CLL co-existing with APL, diagnosed as two separate disease entities based on evidence from molecular testing and immunohistochemistry staining. APL is a hematological emergency. Management of APL, regardless of it being de novo or therapy-related, is the same. Disclosures No relevant conflicts of interest to declare.

SUMMARYCharcot's arthropathy or neuropathic osteoarthropathy is a form of destructive arthropathy associated with decreased sensory innervation. We present an unusual case of a patient referred to oncology clinic for further evaluation of a spindle cell neoplasm of the humerus concerning for malignancy. The mass presented as a lytic lesion associated with a right humerus fracture. This was later diagnosed as Charcot's shoulder secondary to cervical syringomyelia. A detailed case report and discussion are presented here. BACKGROUND

SF1126 is in development in phase I clinical trials as a single agent and interim results have been presented recently in patients with solid tumors (ASCO, 2009, abstract 2558) and multiple myeloma (ASH, 2009, abstract 3879). SF1126 inhibits all four class I phosphatidylinositol 3-kinase (PI3K) isoforms along with other cancer targets such as mTOR, DNA-PK, PIM1, and PLK1. The current ongoing work reported herein describes translational studies providing support for expanding single agent clinical trials (B-cell malignancies such as CLL and mantle cell lymphoma) and combination trials such as with bortezomib in multiple myeloma. Gene expression profiles from myeloma cell lines exposed to SF1126 have identified that HSP-90AA1 is down regulated in common between 3 different myeloma cell lines, while genes for 14-3-3h, cyclin D1, and EIF4BEP1 are commonly downregulated among 2 of the 3 cell lines tested. Genes that are upregulated in common include Bcl-2, Gab2, and CDK2 further supporting the role of cell cycle regulation as a major effect of PI3K inhibition in myeloma cell lines. Additional data on response to SF1126 exposure will be presented in B-cell, T-cell, and mantle cell lymphoma cell lines and primary patient CLL samples. The in vitro combination of SF1126 with dexamethasone (dex) was examined since dex is so commonly used in myeloma patients. Three day proliferation results on RPMI8266 cells (non-responsive to dexamethasone and KRAS mutated) exposed to 5 uM dex plus 5, 10, or 20 uM SF1126 showed a 12, 46, and 78% decrease, respectively, in cell proliferation as measured by the WST staining method versus a no-SF1126-treatment control. The ability of SF1126 to inhibit hematological cancer stem cells was also evaluated. CD138neg cells were isolated from RPMI8226 cell lines as described in the literature (anti-CD138 antibody-conjugated magnetic beads to serially remove CD138+ cells) to give a highly enriched fraction purported to be cancer stem cell ‘like’. SF1126 at 5, 10, or 20 uM demonstrated an 18, 32, and 50% decrease in CD138neg cell proliferation relative to no-treatment controls. These results compared favorably to lenalidomide in the same experiment which showed no decrease in proliferation. These results show for the first time that SF1126 may have activity towards hematological cells characterized as cancer stem cells. In vitro combinations of SF1126 with bortezomib on several myeloma cell lines have previously been shown to exhibit synergistic effects. SF1126 has also previously been shown to have single agent activity in vivo using MM1R (dexamethasone resistant myeloma) xenograft mouse models. The MM1R xenograft mouse model was thus chosen to study SF1126/bortezomib combinations in vivo. Thirty nude mice with similar MM1R tumor sizes were randomly divided into 5 groups of 6 on day 21 post s.c. tumor cell inoculation and treated as follows (all injections were in 100 uL volume administered iv in the tail vein on Tues/Friday 2X/week schedule): 1) saline control; 2) SF1126 at 20 mg/kg; 3) Bortezomib 0.5 mg/kg (1/2 MTD); 4) Bortezomib followed by SF1126 one hour later; 5) SF1126 followed by bortezomib one hour later. Since bortezomib is given iv in the clinic and SF1126 is given as an iv infusion (90 minutes) in the clinic groups 4 and 5 were designed to determine if there is any difference in order of the agents when given in combination. Tumor volume and body weights were assessed through day 42 (3 weeks of treatment). Groups 2, 3, 4, and 5 showed 49, 47, 74, and 85% inhibition of tumor growth relative to the control (group 1). Both of the combination groups 4 and 5 (regardless of order) showed statistical significance compared to control tumor volume (p<0.005). Excised tumor weights showed the same relationship with groups 2, 3, 4, and 5 showing 23, 22, 56, and 66% less tumor weight versus the group 1 control tumor weight with groups 4 and 5 being significant (p<0.01). No treatment related body weight changes were evident and all groups showed increasing body weight gain over the three week treatment period. A second combination study in MM1R xenograft nude mice was performed but dosing was changed to 2X/week using a two weeks on/one week off schedule followed by two weeks more treatment to mimic the clinical schedule using bortezomib (days 1,4,8,11 of a 21 day cycle) and taking into account the clinical administration of SF1126 (twice weekly with no holidays). Twenty nude mice with similar MM1R tumor size were randomly divided into 4 groups of 5 on day 20 post s.c. tumor cell inoculation and treated as follows (all injections were in 100 uL volume administered iv in the tail vein on a 2X/week schedule): 1) saline control; 2) SF1126 at 20 mg/kg; 3) Bortezomib 0.5 mg/kg (1/2 MTD); 4) SF1126 followed by bortezomib one hour later. Tumor volume and body weights were assessed through day 52. This treatment course represented 2 weeks on, one week off, then finally 2 weeks on. Groups 2 and 3 (single agents) showed 70 and 52% inhibition of tumor growth respectively with the combination (group 3) showing the most inhibition at 82% inhibition of tumor growth relative to the control (group 1). Excised tumor weights showed the same relationship with groups 2, 3, and 4 showing 59, 35 and 71% less tumor weight versus the group 1 control tumor weight. No treatment related body weight changes were evident and all groups showed increasing body weight gain over the three week treatment period although the control group showed the largest weight gain. These results indicate in vivo combination effects and help guide the planning of clinical combination studies using SF1126 and bortezomib in multiple myeloma. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-291.

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