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The rhinebothriidean tapeworm family Escherbothriidae has recently been expanded to include the genus Ivanovcestus, species of which parasitise arhynchobatid skates. Similarities in morphology and host associations between Ivanovcestus and Semiorbiseptum - a genus yet to be assigned to one of the families in the order Rhinebothriidea - led us to explore the possibility that Semiorbiseptum might also belong in the Escherbothriidae. Morphological similarities with Scalithrium ivanovae, Scalithrium kirchneri and Rhinebothrium scobinae, all of which also parasitise arhynchobatid skates, raised questions regarding the generic placements of these species. In addition, new collections from the skate Sympterygia brevicaudata revealed two new species that morphologically resemble species of Ivanovcestus. A combination of morphological and molecular data were used to assess the generic placement of the newly discovered species and refine our understanding of the membership of the family Escherbothriidae. Sequence data for the D1-D3 region of the 28S rDNA gene were generated de novo for 14 specimens of 7 rhinebothriidean species and combined with comparable published data to represent all 6 families in the Rhinebothriidea in the analysis. The phylogenetic tree resulting from maximum likelihood analysis strongly supports the inclusion of the genus Semiorbiseptum in the family Escherbothriidae. Our work also suggests that the skate-hosted species previously assigned to Scalithrium and Rhinebothrium are also members of Semiorbiseptum and that Ivanovcestus is a junior synonym of Semiorbiseptum. Six species are transferred to Semiorbiseptum, bringing the total number of species in the genus to ten. The diagnosis of Semiorbiseptum is amended to accommodate the additional species. A second species in the previously monotypic type genus of the family, Escherbothrium, is described. The diagnosis of the Escherbothriidae is amended to include the new and transferred species. This study underscores the importance of integrating morphological and molecular data in bringing resolution to cestode systematics. We believe our findings provide a robust foundation for future research into the evolutionary history and host associations of cestodes within the order Rhinebothriidea and beyond. These also highlight the importance of expanding our understanding of skate-hosted cestodes. [ABSTRACT FROM AUTHOR]

Purpose of Review: The aim of this review is to focus on the recent advances in the molecular knowledge of small cell lung cancer (SCLC) and potential promising new treatment strategies, like targeting the DNA damage pathway, epigenetics, angiogenesis, and oncogenic drivers., Recent Findings: In the last few years, the addition of immunotherapy to chemotherapy has led to significant improvements in clinical outcomes in this complex neoplasia. Nevertheless, the prognosis remains dismal. Recently, numerous genomic alterations have been identified, and they may be useful to classify SCLC into different molecular subtypes (SCLC-A, SCLC-I, SCLC-Y, SCLC-P). SCLC accounts for 10-20% of all lung cancers, most patients have an extensive disease at the diagnosis, and it is characterized by poor prognosis. Despite the progresses in the knowledge of the disease, efficacious targeted treatments are still lacking. In the near future, the molecular characterisation of SCLC will be fundamental to find more effective treatment strategies., (© 2023. The Author(s).)

Aims: In Italy, 12-month survival in the general population between 90 and 94 years old is 26%. In very old patients, the benefit of pacemaker implantation in terms of quality and duration of life is unclear. The aim of our study was to analyse clinical characteristics, outcome and factors associated with survival in patients at least 90 years old at the time of the first pacemaker implant., Methods: Clinical parameters, device characteristics, survival and predictors of outcome in patients at least 90 years old treated with a pacemaker in our centre in 2019-2020 were evaluated., Results: Among the 554 patients undergoing pacemaker implantation in our centre during the study interval, 69 (12%) were at least 90 years old; a complete/advanced atrioventricular block was present in 65%. A cardiological comorbidity (excluding atrial fibrillation) was present in 22 patients (32%). Oncological, pulmonary and neurological comorbidities were present in 12 (17%), 19 (28%) and 32 (46%), respectively. Renal impairment was present in 25 patients (36%). After pacemaker implantation, a pneumothorax developed in two patients and lead dislodgment in one. During follow-up (median 17 months, interquartile range: 13-24), 32 patients died (46%), with a 12-month mortality probability of 24.6%. At multivariate analysis, the presence of oncological (hazard ratio (HR) 5.31; P < 0.001) and neurological (HR 6.44; P < 0.001) comorbidities was associated with mortality. Truncating the outcome at 6 months, renal impairment (HR 8.01; P = 0.003), anticoagulant therapy (HR 8.14; P = 0.003), oncological comorbidities (HR 14.1; P < 0.001) and left ventricular function (5% increase of left ventricular ejection fraction: HR 0.66; P < 0.001) were significantly associated with outcome., Conclusion: At our centre, patients at least 90 years old underwent pacemaker implantation mainly for advanced atrioventricular block. One-year survival was excellent, even better than expected in the general population., (Copyright © 2023 Italian Federation of Cardiology - I.F.C. All rights reserved.)

Hemophilia A is the most common X-linked bleeding disorder affecting more than half-a-million individuals worldwide. Persons with severe hemophilia A have coagulation FVIII levels <1% and experience spontaneous debilitating and life-threatening bleeds. Advances in hemophilia A therapeutics have significantly improved health outcomes, but development of FVIII inhibitory antibodies and breakthrough bleeds during therapy significantly increase patient morbidity and mortality. Here we use sheep fetuses at the human equivalent of 16-18 gestational weeks, and we show that prenatal transplantation of human placental cells (10 7 -10 8 /kg) bioengineered to produce an optimized FVIII protein, results in considerable elevation in plasma FVIII levels that persists for >3 years post-treatment. Cells engraft in major organs, and none of the recipients mount immune responses to either the cells or the FVIII they produce. Thus, these studies attest to the feasibility, immunologic advantage, and safety of treating hemophilia A prior to birth., (© 2023. The Author(s).)

Introduction: Hemophilia A (HA) is the most common X-linked bleeding disorder, occurring in 1 in 5,000 live male births and affecting >1 million individuals worldwide. Although advances in protein-based HA therapeutics have improved health outcomes, current standard-of-care requires infusion 2-3 times per week for life, and 30% of patients develop inhibitors, significantly increasing morbidity and mortality. There are thus unmet medical needs requiring novel approaches to treat HA., Methods: We tested, in a highly translational large animal (sheep) model, whether the unique immunological and biological properties of autologous bone marrow (BM)-derived mesenchymal stromal cells (MSCs) could enable them to serve as cellular delivery vehicles to provide long-term expression of FVIII, avoiding the need for frequent infusions., Results: We show that autologous BM-MSCs can be isolated, transduced with a lentivector to produce high levels of ovine (o)FVIII, extensively expanded, and transplanted into adult animals safely. The transplanted cells engraft in multiple organs, and they stably produce and secrete sufficient quantities of FVIII to yield elevated plasma FVIII levels for at least 15 weeks., Discussion: These studies thus highlight the promise of cellular-based gene delivery approaches for treating HA., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Trevisan, Rodriguez, Medder, Lankford, Combs, Owen, Atala, Porada and Almeida-Porada.)

Background: Immune-checkpoint inhibitors (ICIs) have significantly improved outcome of advanced non-small cell lung cancer (aNSCLC) patients. However, their efficacy remains uncertain in uncommon histologies (UH)., Materials and Methods: Data from ICI treated aNSCLC patients (April,2013-January,2021) in one Institution were retrospectively collected. Univariate and multivariate survival analyses were estimated by Kaplan-Meier and Cox proportional hazards regression model, respectively. Objective response rate (ORR) and disease control rate (DCR) were assessed., Results: Of 375 patients, 79 (21.1%) had UH: 19 (24.1%) sarcomatoid carcinoma, 15 (19.0%) mucinous adenocarcinoma, 10 (12.6%) enteric adenocarcinoma, 8 (10.1%) adenocarcinoma not otherwise specified, 7 (8.9%) large-cell neuroendocrine carcinoma, 6 (7.6%) mixed histology non-adenosquamous, 5 (6.3%) adenosquamous carcinoma, 9 (11.4%) other UH. In UH group, programmed death-ligand 1 (PD-L1) <1%, 1-49%, ≥50% and unknown expression were reported in 27.8%, 22.8%, 31.7% and 17.7% patients respectively and ICI was the second/further-line in the majority of patients. After a median follow-up of 35.64 months (m), median progression-free survival (mPFS) was 2.5 m in UH [95% CI 2.2-2.9 m] versus (vs.) 2.7 m in CH [95% CI 2.3-3.2 m, P-value = .584]; median overall survival (mOS) was 8.8 m [95% CI 4.9-12.6 m] vs. 9.7 m [95% CI 8.0-11.3 m, P-value = .653]. At multivariate analyses only ECOG PS was a confirmed prognostic factor in UH. ORR and DCR were 25.3% and 40.5% in UH vs. 21.6% and 49.5% in CH [P-value = .493 and .155 respectively]., Conclusions: No significant differences were detected between UH and CH groups. Prospective trials are needed to understand ICIs role in UH population., (Copyright © 2021. Published by Elsevier Inc.)

The recognized potential of using mitogenomics in phylogenetics and the more accessible use of high-throughput sequencing (HTS) offer an opportunity to investigate groups of neglected organisms. Here, we leveraged HTS to execute the most comprehensive documentation of mitogenomes for cestodes based on the number of terminals sequenced. We adopted modern approaches to obtain the complete mitogenome sequences of 86 specimens representing five orders of cestodes (three reported for the first time: Phyllobothriidea, "Tetraphyllidea" and Trypanorhyncha). These complete mitogenomes represent an increase of 41% of the mitogenomes available for cestodes (61-147) and an addition of 33% in the representativeness of the cestode orders. The complete mitochondrial genomes are conserved, circular, encoded in the same strand, and transcribed in the same direction, following the pattern observed previously for tapeworms. Their length varies from 13,369 to 13,795 bp, containing 36 genes in total. Except for the Trypanorhyncha specimen, the gene order of the other four cestode orders sequenced here suggests that it could be a synapomorphy for the acetabulate group (with a reversion for taenids). Our results also suggest that no single gene can tell all the evolutionary history contained in the mitogenome. Therefore, cestodes phylogenies based on a single mitochondrial marker may fail to capture their evolutionary history. We predict that such phylogenies would be improved if conducted under a total evidence framework. The characterization of the new mitochondrial genomes is the first step to provide a valuable resource for future studies on the evolutionary relationships of these groups of parasites., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Trevisan, Jacob Machado, Lahr and Marques.)

Microfluidic technology enables recapitulation of organ-level physiology to answer pertinent questions regarding biological systems that otherwise would remain unanswered. We have previously reported on the development of a novel product consisting of human placental cells (PLC) engineered to overexpress a therapeutic factor VIII (FVIII) transgene, mcoET3 (PLC-mcoET3), to treat Hemophilia A (HA). Here, microfluidic devices were manufactured to model the physiological shear stress in liver sinusoids, where infused PLC-mcoET3 are thought to lodge after administration, to help us predict the therapeutic outcome of this novel biological strategy. In addition to the therapeutic transgene, PLC-mcoET3 also constitutively produce endogenous FVIII and von Willebrand factor (vWF), which plays a critical role in FVIII function, immunogenicity, stability, and clearance. While vWF is known to respond to flow by changing conformation, whether and how shear stress affects the production and secretion of vWF and FVIII has not been explored. We demonstrated that exposure of PLC-mcoET3 to physiological levels of shear stress present within the liver sinusoids significantly reduced mRNA levels and secreted FVIII and vWF when compared to static conditions. In contrast, mRNA for the vector-encoded mcoET3 was unaltered by flow. To determine the mechanism responsible for the observed decrease in FVIII and vWF mRNA, PCR arrays were performed to evaluate expression of genes involved in shear mechanosensing pathways. We found that flow conditions led to a significant increase in KLF2, which induces miRNAs that negatively regulate expression of FVIII and vWF, providing a mechanistic explanation for the reduced expression of these proteins in PLC under conditions of flow. In conclusion, microfluidic technology allowed us to unmask novel pathways by which endogenous FVIII and vWF are affected by shear stress, while demonstrating that expression of the therapeutic mcoET3 gene will be maintained in the gene-modified PLCs upon transplantation, irrespective of whether they engraft within sites that expose them to conditions of shear stress., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Trevisan, Morsi, Aleman, Rodriguez, Shields, Meares, Farland, Doering, Spencer, Atala, Skardal, Porada and Almeida-Porada.)

Background: The introduction of immunotherapy has improved the prognosis of patients with Non-Small Cell Lung Cancer (NSCLC). However, data in poor ECOG Performance Status (PS) patients remain scant due to their exclusion from randomized trials., Material and Methods: We analyzed data of patients with advanced NSCLC treated with immunotherapy in two Italian Centers, to evaluate the impact of PS (0-1 vs 2) on disease control rate (DCR), progression free survival (PFS) and overall survival (OS). Chi-square test was used to compare clinical-pathological variables, their impact on survival was evaluated through Cox proportional hazard models., Results: Among 404 patients included, PS was 0 in 137 (33.9 %), 1 in 208 (51.5 %) and 2 in 59 (14.6 %) patients; 143 were female and 90 had squamous NSCLC. Clinical-pathological variables were uniformly distributed except for higher prevalence of liver metastases in patients with poor PS. We found that PS2 patients showed worse outcomes in terms of DCR (21.8 % vs 50.3 %, p = 0.001), PFS [2.0 (95 % CI 1.6-3.0) vs 3.0 (95 % CI 2.7-4.0) months, p < 0.0001] and OS [4.0 (95 % CI 2.8-5.7) vs 13.2 (95 % CI 11.0-15.8) months, p < 0.0001]. PS2 status, negative PDL1 expression and early corticosteroids exposure as well as higher Neutrophil to Lymphocyte Ratio and LDH at baseline were associated with worse outcomes at univariate and multivariable analysis. Subgroup analysis confirmed poor outcomes in PS2 patients with high LDH and concomitant corticosteroid therapies. The incidence of Grade 3/4 adverse events was 11.3 % in PS 0-1 and 10.2 % in PS 2 patients (p = 0.81)., Conclusion: Our data confirm reduced efficacy of immunotherapy in patients with poor PS even though a good safety. Despite PS remains the most powerful independent prognostic factor for NSCLC, LDH levels and steroids exposure could support the decision making in PS2 patients., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Here we describe a new species of RhinebothriumLinton, 1890, from Hypanus guttatus (Bloch and Schneider). Rhinebothrium ramosi n. sp. can be differentiated from all 51 valid species of the genus by having 4-5 testes and uterus that extends throughout the entire length of the proglottid. Only 8 of the above species closely resemble R. ramosi in total length (Rhinebothrium bunburyense, Rhinebothrium chollaense, Rhinebothrium corbatai, Rhinebothrium dasyatidis, Rhinebothrium kruppi, Rhinebothrium lintoni, Rhinebothrium margaritense, and Rhinebothrium reydai). Despite the resemblance, R. bunburyense, R. corbatai, R. dasyatidis, R. lintoni, and R. margaritense can be distinguished from the new species by possessing a larger number of proglottids. The remaining 3 species (R. chollaense, R. kruppi, and R. reydai) overlap in total length and number of proglottids with R. ramosi. However, they can be distinguished from the new species by possessing a single posterior-most bothridial loculus instead of arranged as a pair, as found in the new species. This is the first report of the genus from the coastal waters of Brazil and brings to 52 the number of valid species for this genus. Additionally, we use the patterns of infection and distribution for species of Rhinebothrium to make predictions of expected diversity within the genus, especially for unsurveyed hosts in endemic marine ecoregions of the world., (© American Society of Parasitologists 2020.)

Examination of 4 specimens of the leopard whipray Himantura leoparda, a dasyatid stingray from northern Australia, led to the discovery of 3 new species of Rhinebothrium. Rhinebothrium leopardensis n. sp., Rhinebothrium nandoi n. sp., and Rhinebothrium ruhnkei n. sp. are described, increasing the diversity of the genus to 51 species globally. All 3 new species differ from their congeners in terms of testis number, proglottid number, loculus number, and size. With respect to one another, R. leopardensis n. sp. has bothridia that are weakly constricted at their centers and has a greater number of proglottids than the other 2 species (93-108 vs. 11-15, and 48-78, respectively). Rhinebothrium nandoi n. sp. is the smallest of the 3 species found in H. leoparda (3.6-5 vs. 10-15 mm and 10.1-15.8 mm in total length [TL], respectively) and bears bothridia that are constricted at their centers. Rhinebothrium ruhnkei n. sp. bears bothridia that are conspicuously constricted at their centers and has more testes than R. leopardensis and fewer than R. nandoi (7-10 vs. fewer than 7 and 21-33, respectively). Before this study, 56% (27 of 48) of Rhinebothrium species had been described from the freshwater river systems of South America and the marine waters surrounding South and North America. In contrast, despite the remarkably diverse nature of its batoid fauna, only 19 species were known from the Indo-Pacific region. Our work increases this number to 22, emphasizing the highly underestimated nature of Rhinebothrium diversity in this region of the globe. The discovery of these 3 new species was not unexpected, given the relatively poor status of our current knowledge of the cestode faunas of dasyatid stingrays in the Indo-Pacific region, and given the fact that it is common for a single batoid species to host 2 or more species of Rhinebothrium. Our results suggest that additional work on the cestode faunas of the batoids, especially dasyatids, from the Indo-Pacific region is likely to be highly productive in terms of contributing to the knowledge of Rhinebothrium diversity., (© American Society of Parasitologists 2020.)

Background: Other than the programmed cell death ligand 1 (PD-L1) value, oncologists have only the clinical characteristics of patients with advanced non-small-cell lung cancer (aNSCLC) to determine candidates for immunotherapy. A clinical prognostic score composed of the Eastern Cooperative Oncology Group performance status, sex, histologic type, stage, platinum-based first-line therapy, and response to first-line therapy has categorized 3 prognostic groups for patients undergoing second-line chemotherapy. We sought to validate the same score for patients with aNSCLC treated with second- or further-line immunotherapy., Materials and Methods: We collected data from 2 Italian centers. A score was generated to divide patients into 3 prognostic groups: best, score < 5; intermediate, score 5 to 9; and worst, score > 9. Overall survival (OS) and progression-free survival (PFS) were the endpoints., Results: A total of 347 patients were included for analysis. Their median age was 66 years (range, 30-88 years), most were aged < 70 years (67.5%), 70.7% were men, 79.5% were smokers, and 74.6% had had adenocarcinoma. The Eastern Cooperative Oncology Group performance status was 0 for 23%, 1 for 54.5%, and 2 for 22.5%. Of the 347 patients, 28% were in the best prognosis, 51% in the intermediate, and 21% in the worst prognosis group, respectively. The median OS was 18.0 months for the best, 8.5 months for the intermediate (hazard ratio [HR] vs. best, 1.83; 95% confidence interval [CI], 1.35-2.47; P < .001) and 2.6 months for worst (HR vs. best, 5.77; 95% CI, 3.99-8.33; P < .001) group. The median PFS was 3.4 months for the best, 3.7 months for the intermediate (HR vs. best, 1.35; 95% CI, 1.03-1.77; P = .032), and 1.9 months for the worst (HR vs. best, 2.51; 95% CI, 1.80-3.50; P < .001) group., Conclusions: The prognostic score was able to predict the outcomes of patients with aNSCLC who had received immunotherapy. The worst category showed a dismal life expectancy and probably would not benefit from active systemic therapy. Thus, for these patients, best supportive care could be the best choice., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Several biomarkers have been separately described to select patients for immunotherapy (IO), but few studies integrate these markers. Di Maio, EPSILoN and the plasma microRNA signature classifier (MSC), are three different clinico, biochemical and molecular markers able to independently predict prognosis in non-small cell lung cancer (NSCLC)., Methods: Complete data such as sex, histology, ECOG-PS, stage, smoking status, presence of liver metastasis, LDH and neutrophils-to-lymphocyte ratio were collected to generate Di Maio and EPSILoN. The MSC risk level was prospectively assessed in plasma samples collected at baseline IO. The 3 markers were integrated into the DEMo score system prospectively tested in a cohort of 200 advanced NSCLC patients treated with IO. Endpoints were overall survival (OS), progression-free survival (PFS) and overall response rate (ORR)., Results: DEMo separated patients in 7-risk groups whose median OS had a trend ranging from 29.7 to 1.5 months (P<0.0001). When comparing patients with the lowest (n=29) and the highest (n=35) DEMo scores ORR was 45% and 3%, respectively (P<0.0001). Considering the 53 PD-L1 ≥50% patients, DEMo identified a group of 13 (25%) patients who benefit less from IO in terms of both OS (HR: 8.81; 95% CI: 2.87-20.01) and PFS (HR: 6.82; 95% CI: 2.57-18.10). Twelve out of 111 (11%) patients who most benefit from IO according to OS (HR: 0.21; 95% CI: 0.07-0.62) and PFS (HR: 0.28; 95% CI: 0.12-0.65) were identified by DEMo in the PD-L1 <50% group., Conclusions: The DEMo prognostic score system stratified NSCLC patients treated with IO better than each single marker. The proper use of DEMo according to PD-L1 could improve selection in IO regimens., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at http://dx.doi.org/10.21037/tlcr-20-231). AP reports grants from Italian Association for Cancer Research during the conduct of the study; personal fees from Roche, AstraZeneca and BMS outside the submitted work; in addition, AP has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. CP reports grants from Italian Association for Cancer Research during the conduct of the study; personal fees from BMS and MSD outside the submitted work; in addition, CP has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. GLR reports grants from Italian Association for Cancer Research during the conduct of the study; personal fees from BMS, MSD and AstraZeneca outside the submitted work; in addition, GLR has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. DS reports grants from Italian Association for Cancer Research during the conduct of the study; personal fees from BMS, AstraZeneca and Boehringer Ingelheim outside the submitted work; in addition, DS has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. RF reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, RF has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. GG reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, GG has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. ADT reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, ADT has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. GV reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, GV has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. MB reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, Marta B has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. RL reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, RL has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. BT reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, BT has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. FT reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, FT has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. VT has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. GS reports grants from Italian Association for Cancer Research during the conduct of the study; in addition, GS has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. MCG reports grants from Italian Association for Cancer Research during the conduct of the study; personal fees from Roche, AstraZeneca, BMS, MSD, International GmbH, Boehringer Ingelheim Italia S.P.A, Celgene, Eli Lilly, Ignyta, Incyte, Inivata, MedImmune, Novartis, Pfizer, Roche and Takeda outside the submitted work; in addition, MCG has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. Mattia B reports grants from Italian Ministry of Health during the conduct of the study; in addition, Mattia B has a patent IT1406672 licensed to Gensignia LS, a patent IT1403685 licensed to Gensignia LS, and a patent IT1406866 licensed to Gensignia LS. The other author has no conflicts of interest to declare., (2020 Translational Lung Cancer Research. All rights reserved.)

The delivery of factor VIII (FVIII) through gene and/or cellular platforms has emerged as a promising hemophilia A treatment. Herein, we investigated the suitability of human placental cells (PLCs) as delivery vehicles for FVIII and determined an optimal FVIII transgene to produce/secrete therapeutic FVIII levels from these cells. Using three PLC cell banks we demonstrated that PLCs constitutively secreted low levels of FVIII, suggesting their suitability as a transgenic FVIII production platform. Furthermore, PLCs significantly increased FVIII secretion after transduction with a lentiviral vector (LV) encoding a myeloid codon-optimized bioengineered FVIII containing high-expression elements from porcine FVIII. Importantly, transduced PLCs did not upregulate cellular stress or innate immunity molecules, demonstrating that after transduction and FVIII production/secretion, PLCs retained low immunogenicity and cell stress. When LV encoding five different bioengineered FVIII transgenes were compared for transduction efficiency, FVIII production, and secretion, data showed that PLCs transduced with LV encoding hybrid human/porcine FVIII transgenes secreted substantially higher levels of FVIII than did LV encoding B domain-deleted human FVIII. In addition, data showed that in PLCs, myeloid codon optimization is needed to increase FVIII secretion to therapeutic levels. These studies have identified an optimal combination of FVIII transgene and cell source to achieve clinically meaningful levels of secreted FVIII., (© 2020 The Author(s).)