Prognostic role of lymphocyte to monocyte ratio in patients treated with CAR-T for aggressive lymphoma.

7558 Background: A low absolute lymphocyte to monocyte ratio (ALC/AMC) has been found to predict decreased survival in lymphoma patients receiving chemotherapy and stem cell transplant. We report its clinical significance and additional cellular phenotype changes in patients receiving chimeric antigen receptor T-cell (CART) therapy. Methods: Records were reviewed for patients (pts) who received axicabtagene ciloleucel between 6/2016 and 12/2020. Receiver operator curve was generated using nominal logistic regression to predict CR as best response. Survivals were calculated using Kaplan- Meier method. Blood immune phenotype were assayed by multiparametric flow. Principle component analysis (PCA) was performed using ClusterVis. Results: Low ALC/AMC (≤0.8) prior to lymphodepletion (LD) chemotherapy on day -5 was associated with lower CR rate (AUC=0.68, Table). Our cohort of 81 pts had similar baseline characteristics except that noted in Table. Low ALC/AMC ratio is associated with shorter EFS and OS (EFS: 2.6 vs. 6.4 months, P<0.0001; OS: 5.3 months vs. not reached, P=0.0006), respectively. Prognostic association remained significant in multivariate analysis including ASCT, bridging therapy and CRP. Interestingly, compared to the high ALC/AMC group, the low ALC/AMC group had decreased CD8 Tem, increased CD16+CCR2+ monocytes and increased monocytes’ producing IL12, IL-10, and IL-1β (n=26). Unsupervised PCA identified 3 clusters: 1. Low ALC/AMC, all non-CR; 2. High ALC/AMC, some non-CR; 3. High ALC/AMC, all CR. Compared to cluster 1 and 2, cluster 3 had increased CD4 Tnaive, CD8 Tcm and IL-17 producing CD4 T and NK cells. Conclusions: ALC/AMC is a clinically accessible test that is strongly associated with CAR-T response and survival. Immune characterization revealed that the biologic effect is not just associated with cell ratio. Increased inflammation has been found to negatively impact CAR-T response, with some cytokines known to be from the myeloid lineage. We show that CRP is elevated in the low ALC/AMC group with increased cytokine production by monocytes. In addition, presence of T cell subset and IL-17 producing cells, before LD, are associated with clinical response. Further investigation on optimizing host immunity may help improve clinical outcome with CAR-T.[Table: see text]

Multilayered Porous Titanium-Based 3rd Generation Biomaterial Designed for Endosseous Implants

This work proposes a novel complex multi-layered material consisting of porous titanium as a substrate and a complex coating consisting of a chitosan film engulfing microsphere loaded with growth factors such as BMP2 (bone morphogenic protein 2) and IGF1 (insulin-like growth factor-1). The microspheres were obtained through deposition of dual layers of calcium cross linked pectin–chitosan/pectin polyelectrolyte onto a BSA (bovine serum albumin) gel core. The multilayer was conceived to behave like a 3rd generation biomaterial, by slow delivery of viable growth factors around implants, and to assist the healing of implantation wound and the development of new vital bone. The biologic effect of the delivery of growth factors was studied in vitro, on MSC-CD1 mesenchymal stem cells, and in vivo, on CD1 mice. Proliferation and differentiation of cells were accelerated by growth factors, especially IGF1 for proliferation and BMP2 for differentiation. In vivo tests analyzed histologically and by MicroCT show a more structured tissue around BMP2 samples. The present concept will give the best clinical results if both growth factors are delivered together by a coating film that contains a double population of microcarriers.

Development of a suitable manufacturing process for production of a bioactive recombinant equine chorionic gonadotropin (reCG) in CHO-K1 cells

Equine chorionic gonadotropin (eCG) is a heterodimeric glycoprotein hormone produced by pregnant mares that has been used to improve reproduction activity in different types of livestock. Several strategies to produce the hormone in a recombinant way have been reported; nevertheless, none approach has been able to produce a recombinant eCG (reCG) with significant in vivo bioactivity or in sufficient quantities for commercial purposes. For this reason, the only current product available on the market consists of partially purified preparations from serum of pregnant mares (PMSG). Herein, we describe a highly efficient process based on third-generation lentiviral vectors as delivery method for the production of reCG in suspension CHO-K1 cells, with productivities above 20 IU.106 cell-1.d-1 and 70% purification yields after one purification step. Importantly, reCG not only demonstrated biological activity in bovine cattle but also this bioactivity appeared to be higher than PMSG, since 140 IU of reCG were needed to exert the same biologic effect in an ovulation synchronization protocol compared to 400 IU of PMSG. The results obtained show that the developed strategy represents an attractive option to produce reCG and constitutes an auspicious alternative for the replacement of animals as a source of PMSG.

Effect of ARID1a oncogenic loss of function mutations on overall survival in metastatic breast cancer patients.

e13068 Background: ARID1a (AT Rich Interactive Domain 1A) is part of the SWI/SNF complex, which regulates gene transcription. Its inactivation has been shown to determine resistance to endocrine therapy via enrichment of basal-like gene expression features [Xu G., 2020]. We recently reported that ARID1a lower gene expression might be associated with worse overall survival (OS) [Mariotti V., 2019]. The aim of this study was to analyze the effect of ARID1a mutations on survival in metastatic breast cancer patients (pts). Methods: Breast cancer patients prospectively consented for inclusion in the ORIEN genomic database with known metastatic disease were analyzed using cBioPortal. Predicted biologic effect of mutations from RNASeq data was determined by OncoKB. OS was calculated from initial breast cancer diagnosis to death from any cause. OS analysis using Kaplan Meier and descriptive statistics were performed on SPSS. Results: We identified 644 pts with metastatic breast cancer. Of these, 88 (12.8%) pts harbored an ARID1a mutation. 62 (70.5%) mutations were missense (biologic effect unknown), the remaining 26 (29.5%) were oncogenic loss of function (OLF) changes (frame-shift, deletion, insertion, or nonsense). Median OS was significantly better in patients harboring missense mutations compared to OLF and wild type mutations [median OS 58.2 months (95% CI 44.5-71.8) with missense mutations vs 22.8 months (95% CI 10.8-34.7) with OLF mutations and 27.48 months (95% CI 24.5-30.3) with wild type]. Demographics, tumor features and chemotherapy use were generally equally distributed among the subgroups (table). Conclusions: In our study the median OS was worse in metastatic breast cancer pts harboring OFL ARID1a mutations compared to pts with wild type ARID1a or harboring missense ARID1a mutations. Further studies are warranted to assess how specific ARID1a mutations might affect survival in metastatic breast cancer pts. [Table: see text]

Breastfeeding is associated with reduced risk of multiple sclerosis in males, predominantly among HLA-DRB1*15:01 carriers

Background Breastfeeding as an infant appears protective against later development of some autoimmune diseases, but research into its influence on multiple sclerosis (MS) risk has yielded inconclusive results. Objective We investigated the possible impact of breastfeeding on MS risk. Methods We used two population-based case–control studies comprising 3670 cases and 6737 matched controls. Logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (CI) for association between MS and exposure to prolonged breastfeeding (4 months or longer) versus reduced breastfeeding (less than 4 months). A meta-analysis of case–control studies that assessed the impact of breastfeeding on MS risk among women and men was conducted. Results Prolonged breastfeeding was associated with reduced MS risk among men (OR 0.7, 95% CI 0.5–0.9) but not among women (OR 0.9, 95% CI 0.8–1.1). Among men, a synergistic effect was observed between HLA-DRB1*15:01 carrier status and reduced breastfeeding. Conclusions Findings from the current study add to accumulating evidence that breastfeeding may be a modifiable protective factor for reducing the risk of MS in offspring. When possible, mothers should be supported to breastfeed their infants; however, the mechanism of a sex-specific biologic effect of breastfeeding on MS risk is unclear.

A phase I trial to assess the biologic effect of CBM588 (Clostridium butyricum) in combination with nivolumab plus ipilimumab (nivo/ipi) in patients with metastatic renal cell carcinoma (mRCC).

TPS764 Background: The combination of the immune checkpoint inhibitors (ICIs) nivo/ipi is standard of care for pts with intermediate/poor-risk clear cell RCC. Although data are encouraging, less than 50% of pts will exhibit partial or complete response with nivo/ipi, and approximately 20% of pts will progress through this regimen. Studies have shown that response to ICIs may be modulated by the gut microbiome (Routy et al Science 2018), and that certain gut bacteria (e.g., Bifidobacterium spp) may predispose pts to response (Caitano et al ASCO GU 2017). CBM588 is a strain of Clostridium butyricum, a Gram-positive bacillus which produces short-chain fatty acids. It promotes the growth of commensal gut bacteria like Bifidobacterium, Lactobacillus and Bacteroides spp, and has been shown to enhance Th1 cells and IL-10 production with immunomodulatory effects. Here, we present the study design of an ongoing phase I trial to assess the biologic effect of CBM588 in combination with nivo/ipi in pts with mRCC. Methods: This is a single institution phase I trial. Pts will be randomized to one of two treatment arms. The experimental arm will receive the standard dose/schedule for nivo/ipi (3 mg/kg / 1 mg/kg Q3w for 12w, followed by nivo alone Q4w) plus CBM588 (60 mg PO 3x/d). The control arm will receive nivo/ipi at standard dose/schedule. Key eligibility criteria include intermediate/poor-risk mRCC, no concurrent antibiotic therapy and no prior systemic treatment. The primary objective is to determine the biologic effect of CBM588 + nivo/ipi in the modulation of the gut microbiome, measured by the change in Bifidobacterium spp and Shannon index (a measure of microbiome diversity). Secondary objectives include clinical efficacy (progression free survival by RECIST 1.1) and assessment of systemic immune-modulation effects through the analysis of circulating Tregs, MDSCs, and circulating cytokines/chemokines. Stool samples and serum correlatives will be collected on weeks 1 and 13 of treatment. A two-group t-test with a one-sided type I error of 0.05 will be used to assess the study primary endpoints. Clinical trial information: NCT03829111.

High UBE2T mRNA expression and its prognostic significance in Ovarian cancer: a study based on data mining

Background Ovarian cancer (OC) affects about 22 000 women annually in the US and ranks 5th in cancer deaths, largely due to diagnosed with advanced stage. Epithelial ovarian cancer (EOC) accounts for approximately 90% of all ovarian cancer cases. Our study was to assess the prognostic meaningful of UBE2T expression in OC dependent on data acquired from TCGA and so as to increase further knowledge into the biological pathways involved in OC pathogenesis related to UBE2T. Methods Information on gene expression and comparing clinical data were recognized and downloaded from TCGA. Gene set enrichment analysis (GSEA) created an arranged list of all genes s indicated by their connection with UBE2T expression. Results The scatter plot showed the difference of UBE2T expression between normal and tumor samples ( P <0.01). So as to decide the biological interaction network of UBE2T in OC, we used to tab Network in cBioPortal and the 50 most as often altered neighbor genes of UBE2T were demonstrated utilizing Network and the most frequent alterations were HES1. The GSEA results showed that cell cycle, DNA replication, RNA degradation, some cancers, spliceosome, Huntington’s disease, oxidative phosphorylation are differentially enriched in UBE2T high expression phenotype. Cumulative survive showed that dendritic cell of immune infiltrates statistically significant ( P <0.05) of UBE2T in OC suggesting that dendritic cell significantly affecting the prognosis, it is worth more research and exploration. Conclusion Our study found that the expression of UBE2T was significantly increased in OC patients and associated with several clinical features. UBE2T may be a potentially useful prognostic molecular biomarker of bad survival in OC, while further experimental ought to be performed to demonstrate the biologic effect of UBE2T.

High UBE2T mRNA expression and its prognostic significance in Ovarian cancer: a study based on data mining

Objection Ovarian cancer (OC) affects approximately 22 000 women annually in the US and ranks 5th in cancer deaths, largely due to diagnosed with advanced stage1. Epithelial ovarian cancer (EOC) accounts for approximately 90% of all ovarian cancer cases. Our study was to assess the prognostic meaningful of UBE2T expression in OC dependent on data acquired from TCGA and so as to increase further knowledge into the biological pathways involved in OC pathogenesis related to UBE2T.Method Information on gene expression and comparing clinical data were recognized and downloaded from TCGA. Gene set enrichment analysis (GSEA) created an arranged list of all genes s indicated by their connection with UBE2T expression.Results The scatter plot showed the difference of UBE2T expression between normal and tumor samples (P<0.01). In order to decide the biological interaction network of UBE2T in OC, we applied to tab Network in cBioPortal and the 50 most as often altered neighbor genes of UBE2T were showed utilizing Network and the most frequent alterations were HES1. The GSEA results showed that cell cycle, DNA replication, RNA degradation, some cancers, spliceosome, Huntington’s disease, oxidative phosphorylation are differentially enriched in UBE2T high expression phenotype. Cumulative survive showed that dendritic cell of immune infiltrates statistically significant (P<0.05) of UBE2T in OC indicating that dendritic cell significantly affecting the prognosis, it is worth further research and exploration.Conclusion Our study found that the expression of UBE2T was significantly increased in OC patients and associated with several clinical features. UBE2T may be a potentially useful prognostic molecular biomarker of bad survival in OC, while further experimental ought to be performed to demonstrate the biologic effect of UBE2T.