ALBERT

Description: Background Cancer Cachexia is a metabolic syndrome that can be present even in absence of weight loss and associated with significantly impaired survival. Muscle wasting represents a key-symptom of this syndrome and we recently demonstrated the strong prognosis impact of sarcopenia assessed by computed tomography (CT)-scan in diffuse large B-cell lymphoma (DLBCL) (Lanic et al. Leukemia & Lymphoma 2013). Conversely, the clinical relevance of loss of fat mass (adipopenia) remains unclear. The aim of this study was (i) to investigate the prognostic impact of a multidimensional tool combining a nutritional parameter (albuminemia) and body composition measurements (skeletal muscle and body fat composition) in elderly patients with DLBCL treated by chemotherapy and rituximab (R) (ii) to document the evolution of sarcopenia after immunochemotherapy. Methods This retrospective analysis included 80 DLBCL patients older than 70 years (y) and treated by R-CHOP or R-miniCHOP. Skeletal muscle (SM), visceral (V) and subcutaneal (S) adipose (A) tissues were measured by analysis of stored CT images at the Lumbar vertebrae 3 (L3) level. The surface of the muscular and adipose tissues was selected according to CT Hounsfield unit. Values were normalized for stature to calculate the L3 SM index (LSMI, in cm2/m2), the LVAI and the LSAI and used to define sarcopenia and visceral/subcutaneal adipopenia. Results The characteristics of the patients were as follows: median age = 78 y [70-95]; 36 males; IPI 0-2 = 22, 3-5 = 58; treatment by R-CHOP (n = 45) or R-miniCHOP (n = 35); median body mass index (BMI; in kg/m2) = 23.9. According to the sex-specific defined cut-offs for LSMI (〈 55.8 cm²/m² for men and 38.9 cm²/m² for women), 44 DLBCL patients (55 %, 23 males) were considered as sarcopenic. With a median follow-up of 39 months, the 2y overall survival (OS) in the sarcopenic population was 46% as compared to 84% in the non-sarcopenic group (HR = 3.12; CI95%, 1.66-5.88; p=0.0004). The median LSAI was 76.3 cm2/m2 [10-167] in females and 47.4 cm2/m2[22-100] in males. The median LVSAI was 43.5 cm2/m2[3-141] in females and 50.4 cm2/m2[14-159] in males. Adipopenia, defined by a low LVAI and/or a low LSAI was also highly predictive of the outcome. The 2y OS of the low LVAI population was 48% as compared to 82% for the non-adipopenic group (HR = 2.20; CI95%, 1.19-4.05; p=0.01). The 2y OS in the low LSAI population was 48% as compared to 80% in the non-adipopenic group (HR = 2.28; CI95%, 1.23-4.21; p=0.008). A Three-point cachexia score (CS) including adipopenia, sarcopenia and hypo-albuminemia (defined by an albuminemia 〈 35 g/L) was build and delineates three distinct risk-groups (Figure 1). More importantly the CS remains predictive of the prognosis in a multivariate analysis including BMI (〈 or 〉= 25 kg/m2), age (〈 or 〉= 80y), IPI and gender (HR=2.5; CI95%= 1.14-5.39; p =0.02). LMSI was subsequently reassessed in thirty seven patients during the routine CT scan follow-up [mean = 10 months after pre-treatment CT scan (range 2.8-19.2)]. 15 (40%) patients displayed a 5% decrease of their LSMI, whereas 13 (35%) and 9 (25%) displayed no significant change or increase (〉5%) of the LMSI respectively. Conclusion Our study demonstrates that sarcopenia and adipopenia estimated by CT-scan define cachexia more accurately than BMI or weight loss in elderly DLBCL patients. These factors can be integrated in a cachexia scoring tool which predicts the outcome independently of the BMI and of the IPI. CT scan follow-up indicates that cachexia is a reversible process that should be integrated as part of the therapeutic target in combination with lymphoma treatment. A prospective multicentric trial (registered as NCT01715961/Clinical.gov) is ongoing to validate these anthropometric and nutritional parameters and compare to geriatric assessment scales. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 3681 The prognosis of elderly patients with DLBCL remains particularly poor. The most common explanation involves co-morbidities related to advanced age, which strongly impact chemotherapy feasibility and tolerance. Sarcopenia, defined by the depletion of skeletal muscle, is known to be associated with mortality in individuals with non-malignant diseases but also to be an unfavourable prognosis factor in patients with solid tumors. Its relevance in lymphoma is still unknown. Using a simple and routinely radiological approach, we assessed for the first time the prevalence of sarcopenia and its prognosis value in a population of elderly DLBCL patients. Patients and methods: Sarcopenia was retrospectively determined in 40 DLBCL (median age 78.5 y, range 70–88 y, 19 males), characterized as follow: age adjusted IPI 0–1 = 16, 2–3 = 24; treatment by R-CHOP or R-CHOP-like regimen, n = 39; median comorbidity Charlson index = 3 (range 2–7); mean body mass index (BMI; in kg/m2) = 24.1 (40% with obesity or overweight, no patient classified as under weighted). Muscle mass was measured by analysis of stored CT images obtained for diagnostic purposes before any treatment. A lumbar vertebral landmark (L3) was selected because skeletal muscles in this region correspond to whole-body tissue quantities (Janssen et al. J Appl Physiol 2000). To calculate tissue cross sectional area (cm2), the surface of the muscular tissues was selected according to CT Hounsfield unit (range –29 to 150 for skeletal muscles). This value was normalized for stature to calculate the L3 muscle index (LMI, in cm2/m2). Results: According to the sex-specific cut-offs for LMI defined in solid tumors (55.4 cm2/m2 for men and 38.9 cm2/m2 for women), 19 DLBCL patients (47.5%,10 males) were considered as sarcopenic. Sarcopenic patients, as compared to non sarcopenic patients displayed a similar level of albuminemia, Charlson index, aaIPI, weight loss, BMI, performance status or B symptoms. By contrast the mean age was 81y in the sarcopenic group and 77y in the non sarcopenic group (p=0.003). With a median follow-up of 39 months, the 2y overall survival in the sarcopenic population was 38% as compared to 70% in the non-sarcopenic group (HR = 0.25; CI95%, 0.1–0.70; p=0.01). The prognostic value remains significant in the subgroup of patients younger than 80y (HR=0.12; CI95% 0.03–0.53; p=0.005). Mortality was mainly related to progressive disease in the sarcopenic group. Hypoalbuminemia tend to be correlated to an unfavourable outcome (p=0.06). In multivariate analysis including albuminemia and aaIPI, sarcopenia remains predictive of the outcome (p=0.03). LMI was also calculated after treatment at least 6 months following initial CT scan (n =30, mean interval =11.8 months, range 6.3–19.2). 11 patients (36%) displayed a reduction (〉 5%) and 19 patients (64%) an increase (〉 5%) or a stabilisation of the LMI. Conclusion: Sarcopenia assessed by CT scan appears as a strong prognosis factor in elderly DLBCL patients, more relevant than usual anthropometrical measures or usual prognostic factors. A prospective multicentric study is currently ongoing to validate these results and to determine if the sarcopenic status should be integrated in future strategies to treat elderly patients. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 5073 Population based studies have reported that about 25–35% of DLBCL patients are older than 70 years. It is usually assumed that elderly patients are too frail to receive optimal chemotherapy regarding their frequent co-morbidities, leading to non-manageable treatment toxicity. However clinical trials dedicated to elderly patients demonstrated that a curative approach, based on a dose reduced anthracycline regimen combined with rituximab, is currently realistic in elderly patients. Nutritional parameters may be also crucial in the geriatric setting. Relationships between body compositions, specifically proportions of lean and fat tissues, with cancer incidence and cancer outcomes are of recent interest. In this field, sarcopenia, defined by the depletion of skeletal muscle, is known to be an unfavourable prognosis factor in patients with solid tumours. Its clinical relevance in NHL is still unknown. Taking advantage of the initial staging performed by computed tomography (CT) scan imaging to evaluate lean body mass, we assessed retrospectively the prevalence of sarcopenia and its prognosis value in a population of DLBCL patients older than 70 years treated by chemotherapy and rituximab. Patients and methods This retrospective analysis included 82 patients diagnosed with DLBCL and treated by R-CHOP or R-miniCHOP. Muscle mass was measured by analysis of stored CT images. A lumbar vertebral landmark (L3) was selected because skeletal muscle in this region corresponds to whole-body tissue quantities. To calculate tissue cross sectional area (cm2), the surface of the muscular tissues was selected according to CT Hounsfield unit. This value was normalized for stature to calculate the lumbar L3 skeletal muscle index (LSMI, in cm2/m2). Sex-specific cut-offs for LSMI associated with mortality ascertained by optimum stratification were determined according to “R” for men and women (55. 8 cm2/m2 and 38. 9 cm2/m2 respectively). Results Sarcopenia was determined in 82 DLBCL (median age: 78 y, range 70–95 y, 36 males; age adjusted IPI: 0–1 = 27, 2–3 = 55; mean body mass index (BMI) 24 kg/m2). According to the sex-specific cut-offs for LSMI, 45 DLBCL patients (55%, 24 males) were considered as sarcopenic. Sarcopenic patients, as compared to non sarcopenic patients displayed a similar level of aaIPI. By contrast the mean age was 80y in the sarcopenic group and 77y in the non sarcopenic group (p=0. 006). With a median follow-up of 39 months, the 2y overall survival in the sarcopenic population was 46% as compared to 84% in the non-sarcopenic group (HR = 0. 31; CI95%, 0. 16–0. 57; p=0. 0002). The prognostic value remains significant in the subgroup of patients younger than 80y (n=47) (HR=0. 2; CI95% 0. 07–0. 51; p=0. 0009). Mortality was mainly related to progressive disease in the sarcopenic group. Hypoalbuminemia is related to an unfavourable outcome (p=0. 002). In multivariate analysis including albuminemia, age, treatment and aaIPI, sarcopenia remains predictive of the outcome (p=0. 03). Our results indicate that refine anthropometric measurements based on lean or fat body composition, represent clearly a future field of investigation to optimize rituximab and cytotoxic drugs dose adaptation in a geriatric context. The LSMI, evaluated by routine staging CT scan is a relevant and predictive tool in elderly DLCBL treated by rituximab plus chemotherapy. Sarcopenia defined by this index is related to albuminemia and age but remains predictive of the outcome independently of the aaIPI. A prospective multicentric study is currently on-going to confirm these results and to integrate this anthropometric measurement in a larger comprehensive geriatric assessment. Disclosures: No relevant conflicts of interest to declare.

Description: Overexpression of G protein-coupled receptors (GPCRs) in tumours is widely used to develop GPCR-targeting radioligands for solid tumour imaging in the context of diagnosis and even treatment. The human vasoactive neuropeptide urotensin II (hUII), which shares structural analogies with somatostatin, interacts with a single high affinity GPCR named UT. High expression of UT has been reported in several types of human solid tumours from lung, gut, prostate, or breast, suggesting that UT is a valuable novel target to design radiolabelled hUII analogues for cancer diagnosis. In this study, two original urotensinergic analogues were first conjugated to a DOTA chelator via an aminohexanoic acid (Ahx) hydrocarbon linker and then -hUII and DOTA-urantide, complexed to the radioactive metal indium isotope to successfully lead to radiolabelled DOTA-Ahx-hUII and DOTA-Ahx-urantide. The 111In-DOTA-hUII in human plasma revealed that only 30% of the radioligand was degraded after a 3-h period. DOTA-hUII and DOTA-urantide exhibited similar binding affinities as native peptides and relayed calcium mobilization in HEK293 cells expressing recombinant human UT. DOTA-hUII, not DOTA-urantide, was able to promote UT internalization in UT-expressing HEK293 cells, thus indicating that radiolabelled 111In-DOTA-hUII would allow sufficient retention of radioactivity within tumour cells or radiolabelled DOTA-urantide may lead to a persistent binding on UT at the plasma membrane. The potential of these radioligands as candidates to target UT was investigated in adenocarcinoma. We showed that hUII stimulated the migration and proliferation of both human lung A549 and colorectal DLD-1 adenocarcinoma cell lines endogenously expressing UT. In vivo intravenous injection of 111In-DOTA-hUII in C57BL/6 mice revealed modest organ signals, with important retention in kidney. 111In-DOTA-hUII or 111In-DOTA-urantide were also injected in nude mice bearing heterotopic xenografts of lung A549 cells or colorectal DLD-1 cells both expressing UT. The observed significant renal uptake and low tumour/muscle ratio (around 2.5) suggest fast tracer clearance from the organism. Together, DOTA-hUII and DOTA-urantide were successfully radiolabelled with 111Indium, the first one functioning as a UT agonist and the second one as a UT-biased ligand/antagonist. To allow tumour-specific targeting and prolong body distribution in preclinical models bearing some solid tumours, these radiolabelled urotensinergic analogues should be optimized for being used as potential molecular tools for diagnosis imaging or even treatment tools.

Description: Glioblastomas (GBMs) are the most common primary brain tumors characterized by strong invasiveness and angiogenesis. GBM cells and microenvironment secrete angiogenic factors and also express chemoattractant G protein-coupled receptors (GPCRs) to their advantage. We investigated the role of the vasoactive peptide urotensin II (UII) and its receptor UT on GBM angiogenesis and tested potential ligand/therapeutic options based on this system. On glioma patient samples, the expression of UII and UT increased with the grade with marked expression in the vascular and peri-necrotic mesenchymal hypoxic areas being correlated with vascular density. In vitro human UII stimulated human endothelial HUV-EC-C and hCMEC/D3 cell motility and tubulogenesis. In mouse-transplanted Matrigel sponges, mouse (mUII) and human UII markedly stimulated invasion by macrophages, endothelial, and smooth muscle cells. In U87 GBM xenografts expressing UII and UT in the glial and vascular compartments, UII accelerated tumor development, favored hypoxia and necrosis associated with increased proliferation (Ki67), and induced metalloproteinase (MMP)-2 and -9 expression in Nude mice. UII also promoted a “tortuous” vascular collagen-IV expressing network and integrin expression mainly in the vascular compartment. GBM angiogenesis and integrin αvβ3 were confirmed by in vivo99mTc-RGD tracer imaging and tumoral capture in the non-necrotic area of U87 xenografts in Nude mice. Peptide analogs of UII and UT antagonist were also tested as potential tumor repressor. Urotensin II-related peptide URP inhibited angiogenesis in vitro and failed to attract vascular and inflammatory components in Matrigel in vivo. Interestingly, the UT antagonist/biased ligand urantide and the non-peptide UT antagonist palosuran prevented UII-induced tubulogenesis in vitro and significantly delayed tumor growth in vivo. Urantide drastically prevented endogenous and UII-induced GBM angiogenesis, MMP, and integrin activations, associated with GBM tumoral growth. These findings show that UII induces GBM aggressiveness with necrosis and angiogenesis through integrin activation, a mesenchymal behavior that can be targeted by UT biased ligands/antagonists.