Dual MET and ERBB inhibition overcomes intratumor plasticity in osimertinib-resistant-advanced non-small-cell lung cancer (NSCLC).

BACKGROUND: Third-generation epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) such as osimertinib are the last line of targeted treatment of metastatic non-small-cell lung cancer (NSCLC) EGFR-mutant harboring T790M. Different mechanisms of acquired resistance to third-generation EGFR-TKIs have been proposed. It is therefore crucial to identify new and effective strategies to overcome successive acquired mechanisms of resistance. METHODS: For Amplicon-seq analysis, samples from the index patient (primary and metastasis lesions at different timepoints) as well as the patient-derived orthotopic xenograft tumors corresponding to the different treatment arms were used. All samples were formalin-fixed paraffin-embedded, selected and evaluated by a pathologist. For droplet digital PCR, 20 patients diagnosed with NSCLC at baseline or progression to different lines of TKI therapies were selected. Formalin-fixed paraffin-embedded blocks corresponding to either primary tumor or metastasis specimens were used for analysis. For single-cell analysis, orthotopically grown metastases were dissected from the brain of an athymic nu/nu mouse and cryopreserved at -80°C. RESULTS: In a brain metastasis lesion from a NSCLC patient presenting an EGFR T790M mutation, we detected MET gene amplification after prolonged treatment with osimertinib. Importantly, the combination of capmatinib (c-MET inhibitor) and afatinib (ErbB-1/2/4 inhibitor) completely suppressed tumor growth in mice orthotopically injected with cells derived from this brain metastasis. In those mice treated with capmatinib or afatinib as monotherapy, we observed the emergence of KRAS G12C clones. Single-cell gene expression analyses also revealed intratumor heterogeneity, indicating the presence of a KRAS-driven subclone. We also detected low-frequent KRAS G12C alleles in patients treated with various EGFR-TKIs. CONCLUSION: Acquired resistance to subsequent EGFR-TKI treatment lines in EGFR-mutant lung cancer patients may induce genetic plasticity. We assess the biological insights of tumor heterogeneity in an osimertinib-resistant tumor with acquired MET-amplification and propose new treatment strategies in this situation.

Mutations in NOTCH1 PEST domain orchestrate CCL19-driven homing of chronic lymphocytic leukemia cells by modulating the tumor suppressor gene DUSP22.

Even if NOTCH1 is commonly mutated in chronic lymphocytic leukemia (CLL), its functional impact in the disease remains unclear. Using CRISPR/Cas9-generated Mec-1 cell line models, we show that NOTCH1 regulates growth and homing of CLL cells by dictating expression levels of the tumor suppressor gene DUSP22. Specifically, NOTCH1 affects the methylation of DUSP22 promoter by modulating a nuclear complex, which tunes the activity of DNA methyltransferase 3A (DNMT3A). These effects are enhanced by PEST-domain mutations, which stabilize the molecule and prolong signaling. CLL patients with a NOTCH1-mutated clone showed low levels of DUSP22 and active chemotaxis to CCL19. Lastly, in xenograft models, NOTCH1-mutated cells displayed a unique homing behavior, localizing preferentially to the spleen and brain. These findings connect NOTCH1, DUSP22, and CCL19-driven chemotaxis within a single functional network, suggesting that modulation of the homing process may provide a relevant contribution to the unfavorable prognosis associated with NOTCH1 mutations in CLL.

Bioelectrical impedance vector analysis (BIVA) for the assessment of two-compartment body composition.

This review is directed to define the efficacy of bioelectrical impedance vector analysis (BIVA) for assessing two-compartment body composition. A systematic literature review using MEDLINE database up to 12 February 2014 was performed. The list of papers citing the first description of BIVA, obtained from SCOPUS, and the reference lists of included studies were also searched. Selection criteria included studies comparing the results of BIVA with those of other techniques, and studies analyzing bioelectrical vectors of obese, athletic, cachectic and lean individuals. Thirty articles met the inclusion criteria. The ability of classic BIVA for assessing two-compartment body composition has been mainly evaluated by means of indirect techniques, such as anthropometry and bioelectrical impedance analysis (BIA). Classic BIVA showed a high agreement with body mass index, that can be interpreted in relation to the greater body mass of obese and athletic individuals, whereas the comparison with BIA showed less consistent results, especially in diseased individuals. When a reference method was used, classic BIVA failed to accurately recognize FM% variations, whereas specific BIVA furnished good results. Specific BIVA is a promising alternative to classic BIVA for assessing two-compartment body composition, with potential application in nutritional, sport and geriatric medicine.

Lower lean mass and higher percent fat mass in patients with Alzheimer's disease.

In this study we analyzed body composition in relation to cognitive and functional status, in a cross-sectional sample of patients with Alzheimer's disease (AD). Seventy individuals (27 men, 78.1±6.5years; 43 women, 80.4±5.6years) with mild-moderate stages of AD (clinical dementia ratings 1 and 2) were selected from the Alzheimer Center, SS. Trinità Hospital, ASL 8 of Cagliari (Italy). Cognitive and psycho-functional status was evaluated using mini-mental state examination (MMSE), activities of daily living (ADL) scale, and geriatric depression scale (GDS). Mini-nutritional assessment (MNA) was applied. Anthropometric measurements were taken and body mass index (BMI) was calculated. Body composition was assessed by means of specific bioelectrical impedance vector analysis (BIVA), using the references for the elderly. In comparison with the reference group, patients with AD showed similar BMI and MNA, but peculiar bioelectrical characteristics: lower phase angles and longer vectors (p<0.05). According to specific BIVA, this bioelectrical pattern is indicative of a reduction of lean tissue mass and an increase of percent fat mass (FM%). A more accentuated lean mass reduction (p < 0.05) was observed in women with worse cognitive status and a FM% increase (p < 0.01) in women with worse functional status. In conclusion, patients with AD had lower lean tissue mass and higher percent fat mass than healthy elderly individuals. In women, this pattern was associated with cognitive and functional decline, as indicated by MMSE and ADL values. Specific BIVA showed to be a suitable technique in the elderly, that could enhance BMI and MNA information in the evaluation of nutritional status.

Nutritional and psycho-functional status in elderly patients with Alzheimer's disease.

OBJECTIVES: Analysis of variations of nutritional status in relation to psycho-functional conditions in elderly patients with mild to moderate Alzheimer's disease (AD) by means of bioelectrical impedance vector analysis (BIVA). DESIGN: Cross-sectional study. SETTING: Alzheimer Center, SS. Trinità Hospital, Cagliari (Italy). PARTICIPANTS: 83 free-living patients (29 men, 54 women) with mild-moderate Alzheimer's disease, aged 66 to 96 years, and 91 age-matched controls (37 men and 54 women). MEASUREMENTS: Nutritional status was evaluated by anthropometry (weight, height, waist and upper arm circumferences, triceps skinfold; body mass index, BMI; arm muscle area, AMA); Mini Nutritional Assessment, MNA®; bioelectrical impedance vector analysis, BIVA. Psycho-functional status was assessed by the Mini Mental State Examination (MMSE), Geriatric Depression Scale (GDS), Activities of Daily Living (ADL), and Instrumental Activities of Daily Living (IADL). RESULTS: Compared to the control groups, patients with Alzheimer's disease had a worse psycho-functional and nutritional status. BIVA detected lower body cell mass in Alzheimer's patients with respect to controls (men: T²= 23.4; women: T²=27.3; p<0.01), as well as in the female patients with lower levels of IADL and MMSE (respectively, T²= 8.0; T²=7.4; p<0.05). In patients with AD, a worse psycho-functional status was associated with obesity. CONCLUSION: The psycho-functional decline of patients with AD is related to body composition variations, with a relative increase of fat mass with respect to the muscle component. The BIVA technique distinguished patients from controls and patients with different levels of cognitive decline. Therefore, it is a suitable tool for the screening and monitoring of nutritional status in Alzheimer's disease.