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Advances in treatment have reduced mortality from Hodgkin lymphoma; therefore, greater attention should be focused on minimizing the late effects. A variety of risk-adapted treatment regimens exist that prioritize disease presentation but not patient-specific comorbidities. Herein, we describe a patient with sickle cell disease diagnosed with Hodgkin lymphoma and the considerations made in treatment planning to minimize therapy-related acute toxicity and late effects that overlap with the patient’s preexisting sickle cell disease complications.

Background: Cross-linking of lysine residues in elastic and collagen fibers is a vital process in aortic development. Inhibition of lysyl oxidase by BAPN (β-aminopropionitrile) leads to thoracic aortopathies in mice. Although the renin-angiotensin system contributes to several types of thoracic aortopathies, it remains unclear whether inhibition of the renin-angiotensin system protects against aortopathy caused by the impairment of elastic fiber/collagen crosslinking. Methods: BAPN (0.5% wt/vol) was started in drinking water to induce aortopathies in male C57BL/6J mice at 4 weeks of age for 4 weeks. Five approaches were used to investigate the impact of the renin-angiotensin system. Bulk RNA sequencing was performed to explore potential molecular mechanisms of BAPN-induced thoracic aortopathies. Results: Losartan increased plasma renin concentrations significantly, compared with vehicle-infused mice, indicating effective angiotensin II type 1 receptor inhibition. However, losartan did not suppress BAPN-induced aortic rupture and dilatation. Since losartan is a surmountable inhibitor of the renin-angiotensin system, irbesartan, an insurmountable inhibitor, was also tested. Although increased plasma renin concentrations indicated effective inhibition, irbesartan did not ameliorate aortic rupture and dilatation in BAPN-administered mice. Thus, BAPN-induced thoracic aortopathies were refractory to angiotensin II type 1 receptor blockade. Next, we inhibited angiotensin II production by pharmacological or genetic depletion of AGT (angiotensinogen), the unique precursor of angiotensin II. However, neither suppressed BAPN-induced thoracic aortic rupture and dilatation. Aortic RNA sequencing revealed molecular changes during BAPN administration that were distinct from other types of aortopathies in which angiotensin II type 1 receptor inhibition protects against aneurysm formation. Conclusions: Inhibition of either angiotensin II action or production of the renin-angiotensin system does not attenuate BAPN-induced thoracic aortopathies in mice.

Outcomes for children and adolescents with relapsed and refractory Hodgkin lymphoma (HL) are poor, with approximately 50% of patients experiencing a subsequent relapse. The anti-CD30 antibody-drug conjugate brentuximab vedotin improved progression-free survival (PFS) when used as consolidation after autologous stem cell transplant (ASCT) in adults with high-risk relapsed/refractory HL. Data on brentuximab vedotin as consolidative therapy after ASCT in pediatric patients with HL are extremely limited, with only 11 patients reported in the literature. We performed a retrospective analysis of 67 pediatric patients who received brentuximab vedotin as consolidation therapy following ASCT for the treatment of relapsed/refractory HL to describe the experience of this regimen in the pediatric population. This is the largest cohort reported to date. We found that brentuximab vedotin was well tolerated with a safety profile similar to adult patients. With a median follow up of 37 months, the 3-year PFS was 85%. These data suggest a potential role for the use of brentuximab vedotin as consolidation therapy after ASCT for children with relapsed/refractory Hodgkin lymphoma.

Objective: A cardinal feature of Marfan syndrome is thoracic aortic aneurysm. The contribution of the renin-angiotensin system via AT1aR (Ang II [angiotensin II] receptor type 1a) to thoracic aortic aneurysm progression remains controversial because the beneficial effects of angiotensin receptor blockers have been ascribed to off-target effects. This study used genetic and pharmacological modes of attenuating angiotensin receptor and ligand, respectively, to determine their roles on thoracic aortic aneurysm in mice with fibrillin-1 haploinsufficiency ( Fbn1 C1041G/+ ). Approach and Results: Thoracic aortic aneurysm in Fbn1 C1041G/+ mice was found to be strikingly sexual dimorphic. Males displayed aortic dilation over 12 months while aortic dilation in Fbn1 C1041G/+ females did not differ significantly from wild-type mice. To determine the role of AT1aR, Fbn1 C1041G/+ mice that were either +/+ or −/− for AT1aR were generated. AT1aR deletion reduced expansion of ascending aorta and aortic root diameter from 1 to 12 months of age in males. Medial thickening and elastin fragmentation were attenuated. An antisense oligonucleotide against angiotensinogen was administered to male Fbn1 C1041G/+ mice to determine the effects of Ang II depletion. Antisense oligonucleotide against angiotensinogen administration attenuated dilation of the ascending aorta and aortic root and reduced extracellular remodeling. Aortic transcriptome analyses identified potential targets by which inhibition of the renin-angiotensin system reduced aortic dilation in Fbn1 C1041G/+ mice. Conclusions: Deletion of AT1aR or inhibition of Ang II production exerted similar effects in attenuating pathologies in the proximal thoracic aorta of male Fbn1 C1041G/+ mice. Inhibition of the renin-angiotensin system attenuated dysregulation of genes within the aorta related to pathology of Fbn1 C1041G/+ mice.

Vascular smooth muscle cells (vSMC) exert a critical role in sensing and maintaining vascular integrity. These cells abundantly express the low-density lipoprotein receptor-related protein 1 (LRP1), a large endocytic and signaling receptor that recognizes numerous ligands including ApoE-rich lipoproteins, proteases, and protease-inhibitor complexes. We observed the spontaneous formation of aneurysms in the superior mesenteric artery (SMA) of both male and female mice in which LRP1 was genetically deleted in v SMC (smLRP1-/- mice). Quantitative proteomics revealed elevated abundance of several proteins in smLRP1-/- mice that are known to be induced by angiotensin II (AngII)-mediated signaling, suggesting that this pathway is dysregulated. Administration of losartan, an AngII type I receptor antagonist, or an angiotensinogen antisense oligonucleotide to reduce plasma angiotensinogen concentrations restored the normal SMA phenotype in smLRP1-/- mice and prevented aneurysm formation. Additionally, employing a vascular injury model, we noted excessive vascular remodeling and neointima formation in smLRP1-/- mice that was restored by losartan administration. Together, these findings reveal that LRP1 regulates vascular integrity and remodeling of the SMA by attenuating excessive AngII-mediated signaling.

Advances in treatment have reduced mortality from Hodgkin Lymphoma (HL) and made it so greater attention can be focused on minimizing late effects. A variety of risk-adapted treatment regimens exist that prioritize disease presentation, but not patient-specific comorbidities. Herein we describe a patient with sickle cell disease diagnosed with HL and the considerations made in treatment planning to minimize therapy-related acute toxicity and late effects that overlap with the patient's preexisting sickle cell disease complications.

Atherosclerosis is a leading cause of morbidity and mortality in many countries. Mice are the most frequently used animal model to study the pathogenesis and molecular mechanisms of atherosclerosis. En face analyses of the aorta and cross-sections of the aortic root are the two common modes for quantifying the severity of atherosclerosis in mice. This mini-review introduces these two methods, discusses their pros and cons, and provides suggestions to optimize the quantification of atherosclerosis, thereby enhancing rigor and reproducibility in preclinical research.

A novel Bayesian Monte Carlo integration (BMCI) technique was developed to retrieve geophysical variables from satellite microwave radiometer data in the presence of tropical cyclones. The BMCI technique includes three steps: generating a stochastic database, simulating satellite brightness temperatures using a radiative transfer model, and retrieving geophysical variables such as profiles of temperature, relative humidity, and cloud liquid and ice water content from real observations. The technique also provides uncertainty estimates for each retrieval and can output the error covariance matrix of selected parameters. The measurements from the Advanced Technology Microwave Sounder (ATMS) on board Suomi National Polar-Orbiting Partnership (Suomi NPP) and the Global Precipitation Measurement (GPM) Microwave Imager (GMI) were used as input. A new technique was developed to correct the ATMS and GMI observations for the beam-filling effect, which is due to small-scale variability of precipitation and clouds when compared with the instrument footprint and also the nonlinear relation between the brightness temperature and precipitation. In addition, the assimilation of the BMCI retrievals into the NASA GEOS model is discussed for Hurricane Maria. The results show that assimilating the BMCI retrievals can influence the dynamical features of the cyclone, including a stronger warm core, a symmetric eye, and vertically aligned wind columns. Two possible factors that may limit the impact of the BMCI retrievals include 1) the resolution of the model (about 25 km), which was too coarse to show the potential of the BMCI data in improving the representation of tropical storms in the model forecast, and 2) the data assimilation system not being able to consider vertically correlated observation errors.

Background: The ascending aorta is a common location for aneurysm and dissection. This aortic region is populated by a mosaic of medial and adventitial cells that are embryonically derived from either the second heart field (SHF) or the cardiac neural crest. SHF-derived cells populate areas that coincide with the spatial specificity of thoracic aortopathies. The purpose of this study was to determine whether and how SHF-derived cells contribute to ascending aortopathies. Methods: Ascending aortic pathologies were examined in patients with sporadic thoracic aortopathies and angiotensin II (AngII)–infused mice. Ascending aortas without overt pathology from AngII-infused mice were subjected to mass spectrometry–assisted proteomics and molecular features of SHF-derived cells were determined by single-cell transcriptomic analyses. Genetic deletion of either Lrp1 (low-density lipoprotein receptor–related protein 1) or Tgfbr2 (transforming growth factor–β receptor type 2) in SHF-derived cells was conducted to examine the effect of SHF-derived cells on vascular integrity. Results: Pathologies in human ascending aortic aneurysmal tissues were predominant in outer medial layers and adventitia. This gradient was mimicked in mouse aortas after AngII infusion that was coincident with the distribution of SHF-derived cells. Proteomics indicated that brief AngII infusion before overt pathology occurred evoked downregulation of smooth muscle cell proteins and differential expression of extracellular matrix proteins, including several LRP1 ligands. LRP1 deletion in SHF-derived cells augmented AngII-induced ascending aortic aneurysm and rupture. Single-cell transcriptomic analysis revealed that brief AngII infusion decreased Lrp1 and Tgfbr2 mRNA abundance in SHF-derived cells and induced a unique fibroblast population with low abundance of Tgfbr2 mRNA. SHF-specific Tgfbr2 deletion led to embryonic lethality at E12.5 with dilatation of the outflow tract and retroperitoneal hemorrhage. Integration of proteomic and single-cell transcriptomics results identified PAI1 (plasminogen activator inhibitor 1) as the most increased protein in SHF-derived smooth muscle cells and fibroblasts during AngII infusion. Immunostaining revealed a transmural gradient of PAI1 in both ascending aortas of AngII-infused mice and human ascending aneurysmal aortas that mimicked the gradient of medial and adventitial pathologies. Conclusions: SHF-derived cells exert a critical role in maintaining vascular integrity through LRP1 and transforming growth factor–β signaling associated with increases of aortic PAI1.

Aortic diameter is a standard parameter for defining disease severity of thoracic aortic aneurysms. In mouse studies, aortic diameters can be measured in situ directly, but this approach has a potential confounder of underestimation due to the absence of physiological arterial blood pressure. In the present study, we developed an in situ approach for authentic aortic measurements. Thoracic aortic aneurysms were induced by beta-aminopropionitrile (BAPN, 0.5% wt/vol) administration in 4-week-old male C57BL/6J mice. Ultrasonography was performed to examine aortic dimensions, and mice with thoracic aortic dilatations were terminated subsequently. After saline perfusion through the left ventricle, periaortic tissues were removed to expose thoracic aortas. Optimal cutting temperature (OCT) compound was injected via the left ventricle to maintain aortic patency. In situ aortic images were captured pre- and post-OCT injection. In mice with severe thoracic aortic aneurysms, smaller aortic diameters were observed prior to OCT injection compared to ultrasound measurements, while aortic diameters in situ after OCT were comparable to diameters measured using ultrasound. A telemetry system demonstrated that maximal luminal pressures during 150 ul of OCT injection were 90 mmHg. Immunostaining for CD31 revealed that endothelial cells were preserved in the intima after OCT injection. These results indicate that OCT injection does not cause aortic damages due to excess pressures. In conclusion, in situ imaging with OCT injection provides authentic aortic measurements without overt aortic damage in mice with thoracic aortic aneurysms.

AGT (Angiotensinogen) is the unique substrate of the renin-angiotensin system. Liver is the primary source of circulating AGT. The present study determined whether hepatocyte-derived AGT regulates renal AGT accumulation by injecting ASO (antisense oligonucleotides) targeting hepatocyte-derived AGT (GalNAc AGT ASO) into female cynomolgus monkeys. Hepatocyte-specific inhibition of AGT led to profound reductions of plasma AGT concentrations. AGT protein in S1 and S2 of renal proximal tubules was greatly diminished by GalNAc AGT ASO. Given the similarity between nonhuman primates and human, our findings support the notion that renal AGT is predominantly derived from liver, and liver regulates renal angiotensin II production in humans.Abstract Figure

OBJECTIVE: Completing a college degree is associated with success in employment, financial earnings, and life satisfaction. Mental health difficulties, including attention-deficit/hyperactivity disorder (ADHD), can compromise degree completion. METHOD: We examined 4-year academic performance trajectories of 201 college students with ADHD (97 receiving medication [ADHD-Med], 104 not receiving medication [ADHD-NoMed]) relative to 205 non-ADHD Comparison students. Demographic (e.g., sex, race/ethnicity), psychological (e.g., self-reported depression and anxiety symptoms), and service-related (e.g., receipt of academic support) variables were included as predictors of intercept (i.e., Year 1 performance) and slope (yearly change) of semester GPA, progress toward graduation, and self-reported study skill strategies. RESULTS: College students with ADHD obtained significantly lower GPAs (Hedge’s g = −0.46 and −0.63) and reported less frequent use of study skills strategies (Hedge’s g range from −1.00 to −2.28) than Comparison students. Significantly more Comparison students (59.1%) persisted through eight semesters relative to ADHD-NoMed students (49%). Multiple variables predicted outcomes with parent education, fewer depressive symptoms, better executive functioning, and receipt of high school Section 504 accommodations and college academic support services among the strongest predictors. CONCLUSIONS. Findings suggest support services for students with ADHD should begin prior to college matriculation and focus on improving executive functioning skills and depressive symptoms to increase chances of academic success.

Changes in the concentration of greenhouse gases within the atmosphere lead to changes in radiative fluxes within the atmosphere and at its boundaries. This paper describes an experiment within the...

Abscisic acid (ABA) and gibberellin (GA) are two antagonistic phytohormones that regulate seed germination in response to biotic and abiotic environmental stresses. We demonstrate here that MOTHER OF FT AND TFL1 (MFT), which encodes a phosphatidylethanolamine-binding protein, regulates seed germination via the ABA and GA signaling pathways in Arabidopsis thaliana. MFT is specifically induced in the radical-hypocotyl transition zone of the embryo in response to ABA, and mft loss-of-function mutants show hypersensitivity to ABA in seed germination. In germinating seeds, MFT expression is directly regulated by ABA-INSENSITIVE3 (ABI3) and ABI5, two key transcription factors in ABA signaling pathway. MFT is also upregulated by DELLA proteins in the GA signaling pathway. MFT in turn provides negative feedback regulation of ABA signaling by directly repressing ABI5. We conclude that during seed germination, MFT promotes embryo growth by constituting a negative feedback loop in the ABA signaling pathway.

Graphic AbstractObjectiveA cardinal feature of Marfan syndrome is thoracic aortic aneurysm (TAA). The contribution of ligand-dependent stimulation of angiotensin II receptor type 1a (AT1aR) to TAA progression remains controversial because the beneficial effects of angiotensin receptor blockers have been ascribed to off-target effects. This study used genetic and pharmacologic modes of attenuating angiotensin receptor and ligand, respectively, to determine their roles on TAA in mice with fibrillin-1 haploinsufficiency (Fbn1C1041G/+).Approach and ResultsTAA in Fbn1C1041G/+ mice were determined in both sexes and found to be strikingly sexual dimorphic. Males displayed progressive dilation over 12 months while ascending aortic dilation in Fbn1C1041G/+ females did not differ significantly from wild type mice. To determine the role of AT1aR, Fbn1C1041G/+ mice that were either +/+ or −/− for AT1aR were generated. AT1aR deletion reduced progressive expansion of ascending aorta and aortic root diameter from 1 to 12 months of age in males. Medial thickening and elastin fragmentation were attenuated. An antisense oligonucleotide against angiotensinogen (AGT-ASO) was administered to male Fbn1C1041G/+ mice to determine the effects of angiotensin II depletion. AGT-ASO administration, at doses that markedly reduced plasma AGT concentrations, attenuated progressive dilation of the ascending aorta and aortic root. AGT-ASO also reduced medial thickening and elastin fragmentation.ConclusionsGenetic approaches to delete AT1aR and deplete AngII production exerted similar effects in attenuating pathology in the proximal thoracic aorta of male Fbn1C1041G/+ mice. These data are consistent with ligand (AngII) dependent stimulation of AT1aR being responsible for aortic disease progression.HighlightsProfound sexual dimorphism of aortic disease occurs in Fbn1C1041G/+ mice, with female mice being more resistant and male mice being more susceptible.Inhibition of the AngII-AT1aR axis attenuates aortic pathology in male Fbn1C1041G/+ mice.Antisense oligonucleotides targeting angiotensinogen deplete plasma angiotensinogen and attenuate thoracic aortic aneurysms.

This protocol is used for generation of ascending aortic serial sections for histology. This protocol generates a set of 10 slides covering ~1mm of ascending aortic tissue. Inclusion of the descening aorta provides orientation and a non-TAA control from the same animal.

BackgroundThe ascending aorta is a common location for aneurysm and dissection. This aortic region is populated by a mosaic of medial and adventitial cells that are embryonically derived from either the second heart field (SHF) or the cardiac neural crest. SHF-derived cells populate areas that coincide with the spatial specificity of thoracic aortopathies. The purpose of this study was to determine whether and how SHF-derived cells contribute to ascending aortopathies.MethodsAscending aortic pathologies were examined in patients with sporadic thoracic aortopathies and angiotensin II (AngII)-infused mice. Ascending aortas without overt pathology from AngII-infused mice were subjected to mass spectrometry assisted proteomics, and molecular features of SHF-derived cells were determined by single cell transcriptomic analyses. Genetic deletion of either low-density lipoprotein receptor-related protein 1 (Lrp1) or transforming growth factor-β receptor 2 (Tgfbr2) in SHF- derived cells was conducted to examine the impact of SHF-derived cells on vascular integrity.ResultsPathologies in human ascending aortic aneurysmal tissues were predominant in outer medial layers and adventitia. This gradient was mimicked in mouse aortas following AngII infusion that was coincident with the distribution of SHF-derived cells. Proteomics indicated that brief AngII infusion, prior to overt pathology, evoked downregulation of SMC proteins and differential expression of extracellular matrix proteins, including several LRP1 ligands. LRP1 deletion in SHF-derived cells augmented AngII-induced ascending aortic aneurysm and rupture. Single cell transcriptomic analysis revealed that brief AngII infusion decreased Lrp1 and Tgfbr2 mRNA abundance in SHF-derived cells and induced a unique fibroblast population with low abundance of Tgfbr2 mRNA. SHF-specific Tgfbr2 deletion led to embryonic lethality at E12.5 with dilatation of the outflow tract and retroperitoneal hemorrhage. Integration of proteomic and single cell transcriptomics results identified plasminogen activator inhibitor 1 (PAI1) as the most increased protein in SHF-derived SMCs and fibroblasts during AngII infusion. Immunostaining revealed a transmural gradient of PAI1 in both ascending aortas of AngII-infused mice and human ascending aneurysmal aortas that mimicked the gradient of medial and adventitial pathologies.ConclusionSHF-derived cells exert a critical role in maintaining vascular integrity through LRP1 and TGF-β signaling associated with increases of aortic PAI1.Abstract FigureClinical PerspectiveWhat is new?-SHF-derived SMCs and fibroblasts associate with AngII-induced aortic pathologies.-AngII induces a distinct fibroblast sub-cluster that is less abundant for mRNAs related to major extracellular components and TGFβ ligands and receptors, but more abundant for proliferative genes.-TGFBR2 deletion in SHF-derived cells are embryonic lethal with significant dilatation of the outflow tract in mice.-SHF-specific deletion of LRP1 leads to aortic pathologies in mice, supporting the importance of SHF-derived cells in maintaining ascending aortic wall integrity.What are the clinical implications?-Heterogeneity of the embryonic origins of SMCs and fibroblasts contributes to complex mechanisms of vasculopathy formation, which should be considered when investigating the pathogenesis of thoracic aortopathies.

This study aimed to identify barriers and facilitators to discussing parent mental health within child health care for parents of children with congenital heart disease (CHD). Seventy-nine parents of young children with CHD who received care across 40 hospitals in the United States responded to questions about barriers and facilitators to discussing their mental health with their child’s health care providers. Responses were analyzed using qualitative research methods. Parents described multiple barriers: (1) belief that parent mental health support was outside the care team’s scope of practice, (2) perceived expectation to “stay strong,” (3) fear of negative judgment or repercussion, (4) individual preferences for communication/support, (5) desire to maintain care resources on their child, (6) perceived need to compartmentalize emotions, and (7) negative reactions to past emotional disclosure. Parents also described several facilitators: (1) confidence in the care team’s ability to provide support, (2) intentional efforts by the care team to provide support, (3) naturally extroverted tendencies, and (4) developing personal connections with health care providers. It is important that health care providers normalize the impact of child illness on the family and create an environment in which parents feel comfortable discussing mental health challenges.

This paper describes a forward radiative transfer model and retrieval system (FMRS) for the Tropospheric Water and cloud ICE (TWICE) CubeSat instrument. We use the FMRS to simulate radiances for the TWICE's 14 millimeter- and submillimeter-wavelength channels for a tropical atmospheric state produced by a Weather Research and Forecasting (WRF) model simulation. We also perform simultaneous retrievals of cloud ice particle size, ice water content (IWC), water vapor content (H2O), and temperature from the simulated TWICE radiances using the FMRS. We show that the TWICE instrument is capable of retrieving ice particle size in the range of ~50-1000 μm in mass mean effective diameter with approximately 50% uncertainty. The uncertainties of other retrievals from TWICE are about 1 K for temperature, 50% for IWC and 20% for H2O.

Background:An increased prevalence of obstructive sleep apnoea (OSA) has been suggested in ankylosing spondylitis (AS), but few controlled studies have been performed.Objectives:We thus aimed to study the prevalence of OSA in patients with AS compared to controls and to study if disease-related and non-disease-related factors were determinants of OSA in AS patients.Methods:One hundred and fifty-five patients with AS were included in the Backbone study that investigates severity and comorbidities in AS. Controls were recruited from the Swedish CardioPulmonary bioImage Study (SCAPIS). Participants were asked to be examined with a home sleep-monitoring device during one night’s sleep to evaluate the presence of OSA. For each AS patient, 45-70 years, four controls were matched for sex, age, weight and height. OSA was defined as an apnoea-hypopnea-index ≥5 events/hour.Results:In total, 63/155(40.6%) patients with AS were examined with a home sleep-monitoring device out of which 46 patients were 45-70 years and therefore matched (mean age 57.2±7.5years, 30(65.2%) men) with 179 controls (mean age 57.2±4.5years, 123(68.7%) men). Twenty-two out of 46(47.8%) patients with AS vs. 91/179(50.8%) controls had OSA, p=0.72. No differences measurements evaluating OSA were noted in AS vs. controls. In logistic regression analysis, based on all 63 examined AS-patients, several AS-related variables were associated with OSA but after adjusting for age and sex, only higher age and BMI, remained to be significant determinants of OSA, Table 1.Table 1.Univariable and age- and sex-adjusted logistic regression analyses with obstructive sleep apnoea as dependent variable in 63 patients with ankylosing spondylitis.VariablesUnivariable logistic regression analyses, Odds Ratio (95%CI)PAge- and sex-adjusted logistic regression analyses,Odds Ratio (95%CI)PSex, male1.9(0.6-5.5)0.251.5(0.4-4.8)0.53Age1.1(1.0-1.2)0.0021.1(1.0-1.2)0.002BMI1.4(1.1-1.7)0.0011.6(1.2- 2.2)0.001Duration of symptoms1.1(1.0-1.1)0.0281.0(0.9-1.1)0.79BASMI1.9(1.3-2.9)0.0021.5(0.9 -2.5)0.87BASFI1.4(1.0-2.0)0.0381.3(0.9-2.0)0.88≥1 Syndesmophyte3.9(1.3-12.2)0.0173.0(0.8-11.3)0.10mSASSS1.0(1.0-1.1)0.0471.0(0.98-1.05)0.25Metabolic syndrome4.3(1.5-12.9)0.0081.4(0.3-6.6)0.69Epworth Sleep Scale1.2(1.0-1-3)0.0231.2(1.0-1.4)0.29Conclusion:In this case-control study, patients with AS did not have a higher prevalence of OSA compared to controls. AS patients with OSA had higher BMI and were older compared to patients without OSA.Disclosure of Interests:None declared.

Despite the emergence of YAP as an important regulator of cell proliferation and oncogenesis, a clear picture of the relationship between YAP localization dynamics and downstream transcription is missing. Using live-cell imaging of CRISPR-knockin breast epithelial lines, we show that YAP displays dramatic spatial resets characterized by bulk exit from the nucleus followed by re-entry. These localization resets were upregulated by inhibiting Src-kinase and dampened by Ras transformation. After monolayer wounding, we found that edge-cells exhibited a synchronized localization reset. Lastly, induction of store-operated Ca2+ release and chromatin condensation during mitosis were found to also cause YAP localization resets. We relate these spatiotemporal YAP localization changes to transcriptional outputs from endogenous YAP targets using nascent-transcription reporter-cassette knockins. Maximal transcriptional responses were strictly correlated to the induction of localization resets through targeting Src-kinase, inducing Ca2+ release, and mitosis. By blocking the response to Ca2+ release using a PKC inhibitor, we show that the large transcriptional response is dependent on the ability of YAP to transiently localize to the cytoplasm. We found that Ras transformation, which suppressed YAP localization resets, caused upregulated import and export, reduced overall chromatin binding, and increased target gene expression. Untransformed cells, on the other hand, show relatively high nuclear retention through increased binding and slower nuclear export. This suggests that Ras transformation allows an escape from the compartmentalization-based control shown in untransformed cells. Our results suggest a new perspective on YAP-dependent transcription, revealed through the application of real-time imaging of native YAP dynamics and nascent transcription of YAP target genes. This abstract is also being presented as Poster A21. Citation Format: Matt Franklin, Raja Ghosh, Quanming Shi, Jan Liphardt. Spatial resets modulate YAP-dependent transcription [abstract]. In: Proceedings of the AACR Special Conference on the Hippo Pathway: Signaling, Cancer, and Beyond; 2019 May 8-11; San Diego, CA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(8_Suppl):Abstract nr PR02.

Doxycycline (doxy) is used in treating intracellular and extracellular infections. Liposomal (LE) antibiotics allow low-frequency dosing and extended efficacy compared with standard (STD) formulations. We developed a novel sulfuric acid–loading method for doxycycline liposomes (LE-doxy). We hypothesized that a single s.c. injection of LE-doxy would be detectable in serum for at least 2 weeks at concentrations equal to or better than STD-doxy and would be bactericidal in an in vitro Mycobacterium smegmatis infection of J774A.1 macrophage cells. Liposomes were encapsulated by sulfuric acid gradient loading, and release kinetics were performed in vitro and in vivo. LE-doxy made using 8.25 mg/ml doxycycline loaded for 24 hours achieved 97.77% capture in 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 43.87% in sphingomyelin (sphing). Rats were injected s.c. with 50 mg/kg LE-doxy or 5 mg/kg STD-doxy, and serial blood samples were collected. Pharmacokinetics were analyzed using high-performance liquid chromatography. Liver and injection site skin samples were collected at euthanasia (4 weeks postinjection). Minimal histologic tissue reactions occurred after injection of STD (nonliposomal), DPPC, or sphing-doxy. DPPC-doxy had slightly faster in vitro leakage than sphing liposomes, although both were detectable at 264 hours. The mean residence time for DPPC was the highest (111.78 hours), followed by sphing (56.00 hours) and STD (6.86 hours). DPPC and sphing-doxy were detectable at 0.2 μg/ml in serum at 336 hours postadministration. LE-doxy was not toxic to J774A.1 cells in vitro and produced inhibition of viable Mycobacterium smegmatis at 24 and 48 hours. LE-doxy will require further testing in in vivo infection models.