Chelsea Mayoh

CCI-007, a novel small molecule with cytotoxic activity against infant leukemia with MLL rearrangements

There is an urgent need for the development of less toxic, more selective and targeted therapies for infants with leukemia characterized by translocation of the mixed lineage leukemia (MLL) gene. In this study, we performed a cell-based small molecule library screen on an infant MLL-rearranged (MLL-r) cell line, PER-485, in order to identify selective inhibitors for MLL-r leukemia. After screening initial hits for a cytotoxic effect against a panel of 30 cell lines including MLL-r and MLL wild-type (MLL-wt) leukemia, solid tumours and control cells, small molecule CCI-007 was identified as a compound that selectively and significantly decreased the viability of a subset of MLL-r and related leukemia cell lines with CALM-AF10 and SET-NUP214 translocation. CCI-007 induced a rapid caspase-dependent apoptosis with mitochondrial depolarization within twenty-four hours of treatment. CCI-007 altered the characteristic MLL-r gene expression signature in sensitive cells with downregulation of the expression of HOXA9, MEIS1, CMYC and BCL2, important drivers in MLL-r leukemia, within a few hours of treatment. MLL-r leukemia cells that were resistant to the compound were characterised by significantly higher baseline gene expression levels of MEIS1 and BCL2 in comparison to CCI-007 sensitive MLL-r leukemia cells. In conclusion, we have identified CCI-007 as a novel small molecule that displays rapid toxicity towards a subset of MLL-r, CALM-AF10 and SET-NUP214 leukemia cell lines. Our findings suggest an important new avenue in the development of targeted therapies for these deadly diseases and indicate that different therapeutic strategies might be needed for different subtypes of MLL-r leukemia.

Oral malignant gastrointestinal neuroectodermal tumour with junctional component mimicking mucosal melanoma

Malignant gastrointestinal neuroectodermal tumour (GNET) is a recently characterised rare and aggressive tumour that typically arises in association with the small intestine of adults. We present a novel case of this entity and expand the spectrum of its reported morphological features. The patient was a 5-year-old female, the youngest reported patient affected by the condition, and presented with extra-abdominal disease. The histopathological features included the presence of a junctional component of the palatal tumour, which mimicked mucosal melanoma, a feature that has not been previously reported in GNET. Whole genome and RNA sequencing was performed that demonstrated the EWSR1-ATF1 translocation characteristic of GNET. Knowledge of this entity and its features, together with careful morphological assessment supplemented by judicious immunohistochemical and molecular studies should enable the correct diagnosis to be established.

Delineation of the frequency and boundary of chromosomal copy number variations in paediatric neuroblastoma

ABSTRACT Neuroblastoma, the most common solid tumour in early childhood, is characterized by very frequent chromosomal copy number variations (CNVs). While chromosome 2p amplification, 17q gain, 1p and 11q deletion in human neuroblastoma tissues are well-known, the exact frequencies and boundaries of the chromosomal CNVs have not been delineated. We analysed the publicly available single nucleotide polymorphism (SNP) array data which were originally generated by the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) initiative, defined the frequencies and boundaries of chromosomes 2p11.2 – 2p25.3 amplification, 17q11.1-17q25.3 gain, 1p13.3-1p36.33 deletion and 11q13.3-11q25 deletion in neuroblastoma tissues, and identified chromosome 7q14.1 (Chr7:38254795-38346971) and chromosome 14q11.2 (Chr14:21637401-22024617) deletion in blood and bone marrow samples from neuroblastoma patients, but not in tumour tissues. Kaplan Meier analysis showed that double deletion of Chr7q14.1 and Chr14q11.2 correlated with poor prognosis in MYCN gene amplified neuroblastoma patients. In conclusion, the oncogenes amplified or gained and tumour suppressor genes deleted within the boundaries of chromosomal CNVs in tumour tissues should be studied for their roles in tumourigenesis and as therapeutic targets. Focal deletions of Chr7q14.1 and Chr14q11.2 together in blood and bone marrow samples from neuroblastoma patients can be used as a marker for poorer prognosis and more aggressive therapies.

A risk score including microdeletions improves relapse prediction for standard and medium risk precursor B-cell acute lymphoblastic leukaemia in children

To prevent relapse, high risk paediatric acute lymphoblastic leukaemia (ALL) is treated very intensively. However, most patients who eventually relapse have standard or medium risk ALL with low minimal residual disease (MRD) levels. We analysed recurrent microdeletions and other clinical prognostic factors in a cohort of 475 uniformly treated non‐high risk precursor B‐cell ALL patients with the aim of better predicting relapse and refining risk stratification. Lower relapse‐free survival at 7 years (RFS) was associated with IKZF1 intragenic deletions (P < 0·0001); P2RY8‐CRLF2 gene fusion (P < 0·0004); Day 33 MRD>5 × 10−5 (P < 0·0001) and High National Cancer Institute (NCI) risk (P < 0·0001). We created a predictive model based on a risk score (RS) for deletions, MRD and NCI risk, extending from an RS of 0 (RS0) for patients with no unfavourable factors to RS2 + for patients with 2 or 3 high risk factors. RS0, RS1, and RS2 + groups had RFS of 93%, 78% and 49%, respectively, and overall survival (OS) of 99%, 91% and 71%. The RS provided greater discrimination than MRD‐based risk stratification into standard (89% RFS, 96% OS) and medium risk groups (79% RFS, 91% OS). We conclude that this RS may enable better early therapeutic stratification and thus improve cure rates for childhood ALL.

Brief Report: Potent clinical and radiological response to larotrectinib in TRK fusion-driven high-grade glioma

Genes encoding TRK are oncogenic drivers in multiple tumour types including infantile fibrosarcoma, papillary thyroid cancer and high-grade gliomas (HGG). TRK fusions have a critical role in tumourigenesis in 40% of infant HGG. Here we report the first case of a TRK fusion-driven HGG treated with larotrectinib—the first selective pan-TRK inhibitor in clinical development. This 3-year-old girl had failed multiple therapies including chemotherapy and radiotherapy. Tumour profiling confirmed an ETV6–NTRK3 fusion. Treatment with larotrectinib led to rapid clinical improvement with near total resolution of primary and metastatic lesions on MRI imaging. This is the first report of a TRK fusion glioma successfully treated with a TRK inhibitor.

Abstract 1175: Inhibition of NAMPT as a novel therapeutic strategy for infant leukemia

Novel targeted therapies are urgently needed for infant leukemia as this disease is highly aggressive and refractory to treatment resulting in poor survival rates. The NAD (Nicotinamide Adenine Dinucleotide) producing enzyme NAMPT (Nicotinamide Phosphoribosyltransferase) has been considered an attractive selective target for anti-cancer therapy due to the high dependency of tumor cells on NAD for energy metabolism and activity of NAD-dependent enzymes such as poly-ADP-ribose polymerases (PARPs) and sirtuins that play key roles in cancer cell survival. A newly developed NAMPT inhibitor, OT-82, was initially isolated for its high selective toxicity against a panel of adult leukemia cell lines. Here we investigated NAMPT inhibition as a therapeutic strategy for infant leukemias characterized by rearrangement of the MLL gene (MLL-r), by testing the potency of OT-82 in a panel of preclinical in vitro and in vivo models of MLL-r leukemia that are based on the use of patient-derived xenograft (PDX) cells. OT-82, as a single agent, dramatically reduced the viability of all tested MLL-r leukemia cell lines (n=9) and MLL-r leukemia PDX (n=6) with IC50s ranging from 0.15 to 3.82nM. While the IC50 for OT-82 correlated significantly with the IC50 of other NAMPT inhibitors STF-118804 and FK866, OT-82 was the most potent compound. When combining OT-82 with chemotherapeutic agents currently used to treat infants with leukemia, we observed significant synergy between OT-82 and cytarabine indicating the potential of OT-82 for chemosensitization. Consistent with NAMPT inhibition, OT-82 reduced cytosolic NAD+ levels in MLL-r leukemia cells and inhibited the activity of the NAD-requiring enzymes PARP-1 and SIRT-1, as exemplified by a decrease in PARylated PARP-1 levels and a p53-mediated increase in p21 levels, leading to apoptosis induction. Interestingly, despite the remarkable potency of OT-82 in killing MLL-r leukemia cells, a 25-fold difference in IC50 levels was noted across the cell line panel, with those lines harboring the MLL translocations most prevalent in infants, namely t(4;11) and t(11;19) translocations, being the most sensitive to the compound. A positive correlation was noted between baseline NAMPT mRNA levels and OT-82 IC50 (r=0.7712, P=0.015). Further in vivo testing of OT-82 showed impressive efficacy of the compound in MLL-r leukemia PDX-based animal models (n=6). OT-82 (p.o. 3x week, 40-50 mg/kg, for 3 or 6 weeks) was well tolerated and significantly delayed leukemia progression in 6/6 MLL-r leukemia xenografts with 5/6 achieving objective responses. OT-82 given as monotherapy was as effective as the routinely used triple combination treatment of vincristine, L-asparaginase and dexamethasone. Overall, these results demonstrate that NAMPT inhibition using OT-82 is highly effective against MLL-r leukemia and when combined with current chemotherapies may offer a more selective and potent therapeutic option for infants suffering from this disease. Citation Format: Klaartje Somers, Shiloh Middlemiss, Asel Biktasova, Mawar Karsa, Leanna Cheung, Angelika Kosciolek, Kathryn Evans, Chelsea Mayoh, Ursula R. Kees, Lioubov Korotchkina, Olga B. Chernova, Richard B. Lock, Andrei V. Gudkov, Michelle Haber, Murray D. Norris, Michelle J. Henderson. Inhibition of NAMPT as a novel therapeutic strategy for infant leukemia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1175. doi:10.1158/1538-7445.AM2017-1175