Showing papers by "Oregon Health & Science University published in 2022"

Antibody Response and Variant Cross-Neutralization After SARS-CoV-2 Breakthrough Infection

Abstract: Association of Prior SARS-CoV-2 Infection With Risk of Breakthrough Infection Following mRNA Vaccination in Qatar

WEE1 inhibition induces anti-tumor immunity by activating ERV and the dsRNA pathway.

Abstract: Targeted therapies represent attractive combination partners with immune checkpoint blockade (ICB) to increase the population of patients who benefit or to interdict the emergence of resistance. We demonstrate that targeting WEE1 up-regulates immune signaling through the double-stranded RNA (dsRNA) viral defense pathway with subsequent responsiveness to immune checkpoint blockade even in cGAS/STING-deficient tumors, which is a typical phenotype across multiple cancer types. WEE1 inhibition increases endogenous retroviral elements (ERVs) expression by relieving SETDB1/H3K9me3 repression through down-regulating FOXM1. ERVs trigger dsRNA stress and interferon response, increasing recruitment of anti-tumor T cells with concurrent PD-L1 elevation in multiple tumor models. Furthermore, combining WEE1 inhibition and PD-L1 blockade induced striking tumor regression in a CD8+ T cell-dependent manner. A WEE1 inhibition-induced viral defense signature provides a potentially informative biomarker for patient selection for combination therapy with WEE1 and ICB. WEE1 inhibition stimulates anti-tumor immunity and enhances sensitivity to ICB, providing a rationale for the combination of WEE1 inhibitors and ICB in clinical trials.

An omic and multidimensional spatial atlas from serial biopsies of an evolving metastatic breast cancer

Abstract: •Safe and reliable workflows for multiplatform measurements from single biopsies•Clinical metadata with 11 omic and imaging assays from serial biopsy and blood•Omic, cellular, and structural evolution of metastatic cancer in a single individual•Integrative analyses reveal new potential mechanisms of response and resistance Mechanisms of therapeutic resistance and vulnerability evolve in metastatic cancers as tumor cells and extrinsic microenvironmental influences change during treatment. To support the development of methods for identifying these mechanisms in individual people, here we present an omic and multidimensional spatial (OMS) atlas generated from four serial biopsies of an individual with metastatic breast cancer during 3.5 years of therapy. This resource links detailed, longitudinal clinical metadata that includes treatment times and doses, anatomic imaging, and blood-based response measurements to clinical and exploratory analyses, which includes comprehensive DNA, RNA, and protein profiles; images of multiplexed immunostaining; and 2- and 3-dimensional scanning electron micrographs. These data report aspects of heterogeneity and evolution of the cancer genome, signaling pathways, immune microenvironment, cellular composition and organization, and ultrastructure. We present illustrative examples of how integrative analyses of these data reveal potential mechanisms of response and resistance and suggest novel therapeutic vulnerabilities. Mechanisms of therapeutic resistance and vulnerability evolve in metastatic cancers as tumor cells and extrinsic microenvironmental influences change during treatment. To support the development of methods for identifying these mechanisms in individual people, here we present an omic and multidimensional spatial (OMS) atlas generated from four serial biopsies of an individual with metastatic breast cancer during 3.5 years of therapy. This resource links detailed, longitudinal clinical metadata that includes treatment times and doses, anatomic imaging, and blood-based response measurements to clinical and exploratory analyses, which includes comprehensive DNA, RNA, and protein profiles; images of multiplexed immunostaining; and 2- and 3-dimensional scanning electron micrographs. These data report aspects of heterogeneity and evolution of the cancer genome, signaling pathways, immune microenvironment, cellular composition and organization, and ultrastructure. We present illustrative examples of how integrative analyses of these data reveal potential mechanisms of response and resistance and suggest novel therapeutic vulnerabilities. Precision medicine has led to substantial improvements in clinical outcomes for some individuals with cancer, increasingly through use of analytical procedures that identify people with molecular characteristics associated with an increased likelihood of response.1Von Hoff D.D. Stephenson Jr., J.J. Rosen P. Loesch D.M. Borad M.J. Anthony S. Jameson G. Brown S. Cantafio N. Richards D.A. et al.Pilot study using molecular profiling of patients' tumors to find potential targets and select treatments for their refractory cancers.J. Clin. Oncol. 2010; 28: 4877-4883Crossref PubMed Scopus (437) Google Scholar,2Schwaederle M. Zhao M. Lee J.J. Eggermont A.M. Schilsky R.L. Mendelsohn J. Lazar V. Kurzrock R. Impact of precision medicine in diverse cancers: a meta-analysis of phase II clinical trials.J. Clin. Oncol. 2015; 33: 3817-3825Crossref PubMed Scopus (254) Google Scholar Unfortunately, treatments deployed according to precision medicine principles do not always elicit positive responses, and durable control is achieved for only a subset of individuals with metastatic cancer.3Sanchez N.S. Mills G.B. Mills Shaw K.R. Precision oncology: neither a silver bullet nor a dream.Pharmacogenomics. 2017; 18: 1525-1539Crossref PubMed Scopus (15) Google Scholar We posit that the failure to control individual cancers using biomarker-guided treatments stems in large part from our imperfect understanding of the multitude of resistance mechanisms that drive an individual tumor’s adaptive ability to survive as they evolve under therapy. These mechanisms may involve regulatory networks intrinsic to tumor cells, chemical and mechanical influences from proximal or distal microenvironments, and/or aspects of the immune system. They may vary between individuals with similar biomarkers, across metastases within a person, or among cell subpopulations within a single lesion and may change during treatment. To stimulate and support community-wide investigations of cancer resistance, response, and evolutionary mechanisms in individuals, we present a comprehensive omic and multidimensional spatial (OMS) atlas composed of clinical and research data, response correlates from an affected individuals, and illustrative analytical workflows from a single person with metastatic breast cancer during 3.5 years of treatment. We also illustrate how the atlas can be used to explore spatial and temporal heterogeneity, study tumor evolution, and identify candidate resistance mechanisms and therapeutic vulnerabilities. These studies were carried out in the Serial Measurements of Molecular and Architectural Responses to Therapy (SMMART) program with support from the Human Tumor Atlas Network (HTAN),4Mitri Z.I. Parmar S. Johnson B. Kolodzie A. Keck J.M. Morris M. Guimaraes A.R. Beckett B.R. Borate U. Lopez C.D. et al.Implementing a comprehensive translational oncology platform: from molecular testing to actionability.J. Transl. Med. 2018; 16: 358Crossref PubMed Scopus (22) Google Scholar,5Rozenblatt-Rosen O. Regev A. Oberdoerffer P. Nawy T. Hupalowska A. Rood J.E. Ashenberg O. Cerami E. Coffey R.J. Demir E. et al.The human tumor atlas network: charting tumor transitions across space and time at single-cell resolution.Cell. 2020; 181: 236-249Abstract Full Text Full Text PDF PubMed Scopus (92) Google Scholar through which all data are available in standardized formats. The focus of this OMS atlas is a female individual diagnosed with hormone receptor-positive, HER2-normal, high-OncotypeDx recurrence score,6Paik S. Shak S. Tang G. Kim C. Baker J. Cronin M. Baehner F.L. Walker M.G. Watson D. Park T. et al.A multigene assay to predict recurrence of tamoxifen-treated, node-negative breast cancer.N. Engl. J. Med. 2004; 351: 2817-2826Crossref PubMed Scopus (4546) Google Scholar 0.6-cm right breast ductal carcinoma at the age of 64. She underwent a lumpectomy with intra-operative radiation therapy, followed by treatment with four cycles of adjuvant docetaxel and cyclophosphamide, 2 years of anastrozole, and 5 years of exemestane. Subsequent computed tomography (CT) and fluorodeoxyglucose-positron emission tomography (FDG-PET) scans revealed widespread metastatic disease. The individual was then enrolled in the SMMART program (Figure 1). Management decisions were made by the treating physician based on all clinical information plus input from a multidisciplinary tumor board (STAR Methods). This led to four treatment phases over a 3.5-year period (Figure 2A). Temporary tumor control was achieved in the first three phases, with a new phase of therapy beginning at signs of progression. Toxicity of the combination therapies was effectively managed through supportive medication and dose reduction (Figure S2A). Standard toxicity-related blood chemistries were monitored, including absolute neutrophil and platelet counts and liver function tests (Figures S2B–S2D; Table S1).Figure 2Timeline of clinical treatment and response metricsShow full caption(A) Treatment schedule and biopsy timing (red stars) over four phases of treatment (green, orange, blue, and pink areas). The timeline is sectioned into 28-day months. The duration and relative dose for each drug is indicated by the extent and width of a horizontal bar. Drug continuation after the end of phase 4 is indicated by a right arrow.(B) Clinically reported serum levels of tumor protein biomarkers. CEA values were multiplied by 5 to ease visualization.(C) RECIST 1.1 assessment of tumor response (orange stars) indicating partial response (PR), progressive disease (PD), or stable disease (SD). Shown are longitudinal tracking and variation in the longest-axis size of 16 representative metastatic lesions measured from serial CT images. Targets of metastatic biopsies are bolded and marked with stars. Circles represent FDG-PET imaging results, colored and centered on the lines of their corresponding lesion at interpolated lesion sizes. The diameter of each circle is proportional to the background-normalized maximum standardized uptake value (SUVmax).View Large Image Figure ViewerDownload Hi-res image Download (PPT) (A) Treatment schedule and biopsy timing (red stars) over four phases of treatment (green, orange, blue, and pink areas). The timeline is sectioned into 28-day months. The duration and relative dose for each drug is indicated by the extent and width of a horizontal bar. Drug continuation after the end of phase 4 is indicated by a right arrow. (B) Clinically reported serum levels of tumor protein biomarkers. CEA values were multiplied by 5 to ease visualization. (C) RECIST 1.1 assessment of tumor response (orange stars) indicating partial response (PR), progressive disease (PD), or stable disease (SD). Shown are longitudinal tracking and variation in the longest-axis size of 16 representative metastatic lesions measured from serial CT images. Targets of metastatic biopsies are bolded and marked with stars. Circles represent FDG-PET imaging results, colored and centered on the lines of their corresponding lesion at interpolated lesion sizes. The diameter of each circle is proportional to the background-normalized maximum standardized uptake value (SUVmax). The clinical metadata in Table S1 link detailed treatment doses and timelines (Figure 2A) to tumor response metrics. Serum levels of the tumor protein biomarkers carcinoembryonic antigen (CEA), cancer antigen 15-3 (CA 15-3), and cancer antigen 27-29 (CA 27-29) were routinely measured to monitor treatment response (Figure 2B; Table S1). Increasing biomarker levels are concerning for underlying progression, but National Comprehensive Cancer Network (NCCN) guidelines do not recommend changing therapies solely based on blood biomarkers.7National Comprehensive Cancer NetworkBreast cancer (Version 8.2021).https://www.nccn.org/professionals/physician_gls/pdf/breast.pdfGoogle Scholar Biomarker measurements were thus complemented by periodic CT and FDG-PET imaging, with response evaluated using response evaluation criteria in solid tumors (RECIST) 1.1 criteria8Eisenhauer E.A. Therasse P. Bogaerts J. Schwartz L.H. Sargent D. Ford R. Dancey J. Arbuck S. Gwyther S. Mooney M. et al.New response evaluation criteria in solid tumours: revised RECIST guideline (version 1.1).Eur. J. Cancer. 2009; 45: 228-247Abstract Full Text Full Text PDF PubMed Scopus (15771) Google Scholar (Figure 2C; Table S1). Representative computed tomography (CT), fluorodeoxyglucose-positron emission tomography (FDG-PET), and ultrasound images highlight disease burden at key time points (Figure S1). Biospecimens collected for analysis include serial blood samples, a primary breast tumor (PT), a liver biopsy taken immediately prior to phase 1 (Bx1), a biopsy of a different liver lesion taken at the end of phase 1 (Bx2), a bone lesion biopsy taken at the end of phase 3 (Bx3), and a biopsy of a third liver lesion taken at the end of phase 4 (Bx4; Figure 2A). Importantly, Bx2–Bx4 were acquired from metastatic lesions explicitly identified on serial CT and/or FDG-PET imaging as progressing near the end of each respective treatment phase (Figures 2A and S1A–S1I). These biospecimens were analyzed using 11 distinct omic and multiscale spatial imaging workflows to generate this OMS atlas (Figure 1). Targeted DNA sequencing (GeneTrails solid tumor panel, all biopsies), whole-exome sequencing (WES; primary tumor and Bx1–Bx4), and low-pass whole genome sequencing (LP-WGS; Bx3 and Bx4) were used to identify somatic genomic alterations, including single-nucleotide variants (SNVs), insertions or deletions (indels), and copy number changes (Figures 3A and S3A). Ubiquitous alterations included amplification of the CDK4/6 regulatory partner cyclin D1 (CCND1; Figure S3B). Other biologically and clinically relevant alterations were private to a single biopsy. For example, Bx2 contained a hotspot PIK3CA mutation (p.E542K; GenBank: NM_006218:c.1624G>A)9Dogruluk T. Tsang Y.H. Espitia M. Chen F. Chen T. Chong Z. Appadurai V. Dogruluk A. Eterovic A.K. Bonnen P.E. et al.Identification of variant-specific functions of PIK3CA by rapid phenotyping of rare mutations.Cancer Res. 2015; 75: 5341-5354Crossref PubMed Scopus (88) Google Scholar,10Ng P.K. Li J. Jeong K.J. Shao S. Chen H. Tsang Y.H. Sengupta S. Wang Z. Bhavana V.H. Tran R. et al.Systematic functional annotation of somatic mutations in cancer.Cancer Cell. 2018; 33: 450-462.e410Abstract Full Text Full Text PDF PubMed Scopus (96) Google Scholar that was absent from other samples and was taken from a liver lesion that increased in size during treatment with drugs that target aspects of phosphatidylinositol 3-kinase (PI3K) signaling (Figures 2A and 2C). Bx3 and Bx4 both harbored similar amplified regions on chromosome 18 that were not detected in prior biopsies or the primary tumor; Bx3 had 8 copies of a 2.3-Mb region, and Bx4 had 14 copies of a 0.7-Mb region (Figures 3B and S3B). These amplicons contained the genes for thymidylate synthetase (TYMS) and the SRC family tyrosine kinase YES1 and were accompanied by increased TYMS and YES1 RNA relative to Bx2 (TYMS: B×3 = 6.8×, B×4 = 7.2×; YES1: B×3 = 2.0×, B×4 = 4.0×). Importantly, both biopsies were acquired after treatment with the TYMS inhibitor capecitabine (Figures 2A, 2C, and S1G). A phylogenetic analysis revealed that Bx3 diverged from the primary tumor at an earlier evolutionary stage than Bx1, Bx2, or Bx4 (Figure 3B) but was only detected on FDG-PET imaging 1 month before the biopsy occurred (Figures S1A, S1F, and S1G). WES of circulating tumor DNA (ctDNA) from blood collected immediately prior to Bx1 (ctDNA1) and 23 days after Bx2 (ctDNA2) showed that ctDNA1 carried mutations identified previously as private to Bx2, Bx3, or Bx4, whereas ctDNA2 had mutations that were private to each of the four biopsies (Figure S3A). Thus, at least some of the genomic features detected in later biopsies were present before initiation of treatment. Tumor mutational burden (TMB) was assessed for the primary tumor and Bx1–Bx4 because a TMB of 10 or more mutations per megabase (mut/Mb) has been associated with a positive response to immune checkpoint blockade.11O'Meara T.A. Tolaney S.M. Tumor mutational burden as a predictor of immunotherapy response in breast cancer.Oncotarget. 2021; 12: 394-400Crossref PubMed Google Scholar The TMB was low overall (1.2–5.2 mut/Mb), but we identified 1,271 unique neoepitopes (158–687 neoepitopes per biopsy) predicted to bind to at least one major histocompatibility complex (MHC) allele with an affinity of less than 500 nM (Table S2). Human leukocyte antigen (HLA) subtypes were stable across all biopsies, and no loss of heterozygosity was observed. Notably, 68 neoepitopes that might serve as targets for a personalized cancer vaccine were present in the primary tumor and all four biopsies (Figure S4A).12McGranahan N. Furness A.J. Rosenthal R. Ramskov S. Lyngaa R. Saini S.K. Jamal-Hanjani M. Wilson G.A. Birkbak N.J. Hiley C.T. et al.Clonal neoantigens elicit T cell immunoreactivity and sensitivity to immune checkpoint blockade.Science. 2016; 351: 1463-1469Crossref PubMed Scopus (1655) Google Scholar,13Blass E. Ott P.A. Advances in the development of personalized neoantigen-based therapeutic cancer vaccines.Nat. Rev. Clin. Oncol. 2021; 18: 215-229Crossref PubMed Scopus (65) Google Scholar Dual index degenerate adaptor sequencing (DIDA-seq)14Butler T.M. Boniface C.T. Johnson-Camacho K. Tabatabaei S. Melendez D. Kelley T. Gray J. Corless C.L. Spellman P.T. Circulating tumor DNA dynamics using patient-customized assays are associated with outcome in neoadjuvantly treated breast cancer.Cold Spring Harb. Mol. Case Stud. 2019; 5: a003772Crossref PubMed Scopus (12) Google Scholar was performed, using a panel of 53 SNVs present in the individual’s primary tumor, Bx1, and/or Bx2 to assess ctDNA levels from serial plasma samples collected over the first 32 months of treatment (Table S2). The average variant allele frequency (VAF) of the SNV panel remained below 0.3% of total cell free DNA during this period, with the exception of two transient increases (Figure S2E). The first occurred immediately prior to Bx2 (Figure S2E), coincident with rising CA 15-3 and CA 27-29 levels (Figure 2B), followed by progressive disease (PD; Figure 2C). The increase in ctDNA VAF was greatest for mutations shared by the primary tumor, Bx1, and Bx2 (30% VAF) compared with those private to the metastases (Bx1 and Bx2, 3.1%; Bx1, 0.05%; Bx2, 1.3%). A second ctDNA increase occurred after palliative radiation therapy to spinal lesions at C2–C5. Interestingly, the VAFs of all SNV groups in the panel increased at this time, including those private to Bx1 and Bx2 liver lesions. Signaling and pathway activities were calculated from whole-transcriptome sequencing (RNA sequencing [RNA-seq]). Classification using the PAM50 subtype gene signature15Parker J.S. Mullins M. Cheang M.C. Leung S. Voduc D. Vickery T. Davies S. Fauron C. He X. Hu Z. et al.Supervised risk predictor of breast cancer based on intrinsic subtypes.J. Clin. Oncol. 2009; 27: 1160-1167Crossref PubMed Scopus (2662) Google Scholar showed liver biopsies Bx1, Bx2, and Bx4 to be luminal A, whereas the bone biopsy Bx3 was luminal B (Figure S3C). Table S2 summarizes RNA transcript levels and pathway activity estimates for Bx1–Bx4 relative to breast cancers in The Cancer Genome Atlas (TCGA-BRCA)16Cancer Genome Atlas NetworkComprehensive molecular portraits of human breast tumours.Nature. 2012; 490: 61-70Crossref PubMed Scopus (7484) Google Scholar and gene set variation analysis (GSVA) of enriched Molecular Signatures Database (MSigDB) cancer hallmarks relative to TCGA-BRCA luminal samples.17Hanzelmann S. Castelo R. Guinney J. GSVA: gene set variation analysis for microarray and RNA-seq data.BMC Bioinformatics. 2013; 14: 7Crossref PubMed Scopus (2171) Google Scholar,18Liberzon A. Birger C. Thorvaldsdottir H. Ghandi M. Mesirov J.P. Tamayo P. The Molecular Signatures Database (MSigDB) hallmark gene set collection.Cell Syst. 2015; 1: 417-425Abstract Full Text Full Text PDF PubMed Scopus (2519) Google Scholar Proliferation, Immune, and Signaling were the most variable (MSigDB) Hallmark Process categories across the biopsies (Figure 3C). Notably, Bx2 harboring the PIK3CA p.E542K mutation had reduced “PI3K/AKT/mTOR” compared with Bx1, although that gene set was still increased relative to TCGA-BRCA samples. Protein and phosphoprotein abundances were measured in Bx1, Bx2, and Bx4 using reverse-phase protein arrays (RPPAs), and proteomic pathway signatures were compared with TCGA-BRCA (Figures 3D and 3E).19Chen M.M. Li J. Wang Y. Akbani R. Lu Y. Mills G.B. Liang H. TCPA v3.0: an integrative platform to explore the pan-cancer analysis of functional proteomic data.Mol. Cell Proteomics. 2019; 18: S15-S25Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar, 20Labrie M. Fang Y. Kendsersky N.D. Li J. Liang H. Westin S.N. Mitri Z. Mills G.B. Using reverse phase protein array (RPPA) to identify and target adaptive resistance.Adv. Exp. Med. Biol. 2019; 1188: 251-266Crossref PubMed Scopus (2) Google Scholar, 21Tibes R. Qiu Y. Lu Y. Hennessy B. Andreeff M. Mills G.B. Kornblau S.M. Reverse phase protein array: validation of a novel proteomic technology and utility for analysis of primary leukemia specimens and hematopoietic stem cells.Mol. Cancer Ther. 2006; 5: 2512-2521Crossref PubMed Scopus (490) Google Scholar, 22Labrie M. Kim T.B. Ju Z. Lee S. Zhao W. Fang Y. Lu Y. Chen K. Ramirez P. Frumovitz M. et al.Adaptive responses in a PARP inhibitor window of opportunity trial illustrate limited functional interlesional heterogeneity and potential combination therapy options.Oncotarget. 2019; 10: 3533-3546Crossref PubMed Scopus (8) Google Scholar Aspects of hormone signaling varied across biopsies. Estrogen receptor (ER) protein levels from RPPA and clinical immunohistochemistry (IHC) were high in all three biopsies (Table S1). The protein pathways “hormone signaling” and “hormone receptor” were higher in Bx2 (Figure 3D), whereas the GSVA RNA hallmarks “estrogen early” and “estrogen late” (Figure 3C) showed little change, an intriguing finding because protein levels of the hormone-regulated transcription factors ER, GATA3, and adrenergic receptor (AR) were increased in Bx2 relative to Bx1 after phase 1 treatments (Figure 3E). Bx4, taken after phase 4 treatment without hormone suppression, showed continued elevation of the “hormone receptor” pathway and ER and AR protein levels relative to Bx1. However, GATA3 protein levels, the “hormone signaling” protein pathway, and the “estrogen early” and “estrogen late” GSVA hallmarks were downregulated. PI3K/AKT/mTOR pathway signaling from RPPA was generally similar across all biopsies, even though Bx2 was collected after treatment with the mTORC1 inhibitor everolimus and contained the hotspot mutation PIK3CA p.E542K (Figure 3D). Individual protein levels within these pathways varied but did not result in changes in overall signaling. For example, Bx2 showed decreased mTORC1 complex activity based on decreased S6 phosphorylation at S235/236 and S240/244 (0.7× and 0.1× versus Bx1) but increased activity downstream of mTORC2, including increased phosphorylation of AKT (S473: 2.7× versus Bx1) and its substrates GSK3A/B (S21/S9: 1.7× versus Bx1), TSC2 (T1462: 1.4× versus Bx1), and MDM2 (S166: 1.8× versus Bx1; Figure 3E). Likewise, Bx4 showed increased phosphorylation of AKT at S473 (2.7× versus Bx1, 1.0× versus Bx2) and NDRG1 (T346: 1.8× versus Bx1, 1.6× versus Bx2) but without an accompanying increase in AKT or mTORC1 substrate phosphorylation. We also used the clinical Intracellular Signaling Protein Panel (ISPP) to quantitate phosphoproteins and total proteins in Bx2–Bx4 (Figure 3F; Table S2).23Lee J. Geiss G.K. Demirkan G. Vellano C.P. Filanoski B. Lu Y. Ju Z. Yu S. Guo H. Bogatzki L.Y. et al.Implementation of a multiplex and quantitative proteomics platform for assessing protein lysates using DNA-barcoded antibodies.Mol. Cell Proteomics. 2018; 17: 1245-1258Abstract Full Text Full Text PDF PubMed Scopus (13) Google Scholar ISPP showed that Bx2 had the highest AKT phosphorylation and lowest S6 phosphorylation relative to the other biopsies, consistent with RPPA results, whereas p-ERK, p-cRAF, and p-MEK were elevated in Bx3 and increased relative to other biopsies. A transcriptional regulator analysis using a molecular interactions network derived from Pathway Commons24Cerami E.G. Gross B.E. Demir E. Rodchenkov I. Babur O. Anwar N. Schultz N. Bader G.D. Sander C. Pathway Commons, a web resource for biological pathway data.Nucleic Acids Res. 2011; 39: D685-D690Crossref PubMed Scopus (720) Google Scholar was used to infer regulator protein activity from the gene expression data. Integrative analysis of the longitudinal changes in proteomics, gene expression, and transcriptional regulator scores between Bx1 and Bx2 was also performed using CausalPath (Figure S3D).25Babur Ö. Luna A. Korkut A. Durupinar F. Siper M.C. Dogrusoz U. Vaca Jacome A.S. Peckner R. Christianson K.E. Jaffe J.D. et al.Causal interactions from proteomic profiles: molecular data meet pathway knowledge.Patterns. 2021; 2: 100257Abstract Full Text Full Text PDF PubMed Scopus (0) Google Scholar These analyses showed strong inhibition of mTOR regulator activity (Bx2 5.1× > Bx1). Activities of multiple JAK-STAT family proteins were increased, including JAK2 (Bx2 1.8× > Bx1), phospho-STAT3 (Y705: Bx2 1.6× > Bx1), and STAT5 (Bx2 3.2× > Bx1), which, together with the oncoprotein mucin 1 (MUC1; protein Bx2 27.1× > Bx1; regulator Bx2 +3.15 versus Bx1) constitute a known feedforward loop whereby MUC1 binds STAT3 to facilitate its phosphorylation by JAK1.26Ahmad R. Rajabi H. Kosugi M. Joshi M.D. Alam M. Vasir B. Kawano T. Kharbanda S. Kufe D. MUC1-C oncoprotein promotes STAT3 activation in an autoinductive regulatory loop.Sci. Signal. 2011; 4: ra9Crossref PubMed Scopus (71) Google Scholar These observations are reinforced by elevation in “IL6/JAK/STAT3” and “IL2/STAT5” signaling from GSVA (Figure 3C). This analysis also highlighted decreases in MYC and E2F regulator activity and E2F1 total protein, consistent with decreased enrichment of “MYC targets” and “E2F1 targets” in GSVA. These analyses provide a view of the interaction dynamics of cell cycle control networks, with decreases in the expression of cell cycle progression genes (CCNB1, CDK4, CDK1, CCNE2, CCND3, and PLK1) balanced by a sharp decrease in cell cycle inhibitor genes (CDKN1A, CDKN1B, and CDKN2A), leaving RB1 phosphorylation unchanged in Bx2. Changes in composition and functionality of lymphoid and myeloid lineage immune cells were assessed in the primary tumor and Bx1–Bx4 using multiplex IHC (mIHC),27Banik G. Betts C.B. Liudahl S.M. Sivagnanam S. Kawashima R. Cotechini T. Larson W. Goecks J. Pai S.I. Clayburgh D.R. et al.High-dimensional multiplexed immunohistochemical characterization of immune contexture in human cancers.Methods Enzymol. 2020; 635: 1-20Crossref PubMed Scopus (14) Google Scholar, 28Tsujikawa T. Kumar S. Borkar R.N. Azimi V. Thibault G. Chang Y.H. Balter A. Kawashima R. Choe G. Sauer D. et al.Quantitative multiplex immunohistochemistry reveals myeloid-inflamed tumor-immune complexity associated with poor prognosis.Cell Rep. 2017; 19: 203-217Abstract Full Text Full Text PDF PubMed Scopus (240) Google Scholar, 29Tsujikawa T. Thibault G. Azimi V. Sivagnanam S. Banik G. Means C. Kawashima R. Clayburgh D.R. Gray J.W. Coussens L.M. et al.Robust cell detection and segmentation for image cytometry reveal Th17 cell heterogeneity.Cytometry A. 2019; 95: 389-398Crossref PubMed Scopus (9) Google Scholar noting possible discordance in Bx3 because of its bone origin (Figures 4 and S4B–S4E; Table S2). Total immune cell infiltration, as indicated by the percentage of CD45+ cells, was lowest in Bx2 (2.1%) but comparable between the PT (7.2%), Bx1 (9.3%), and Bx4 (10.0%; Figures 4A and 4B). Myelomonocytic cells (macrophages and monocytes) comprised the dominant CD45+ leukocyte lineage subgroup in the PT (65.8%; Figure 4C, green and brown), Bx1 (48.3%), and Bx2 (82.4%) and were reduced in Bx4 (6.1%). Analysis of the myeloid lineage revealed that the fraction of immature dendritic cells was higher in Bx1 (0.2%; Figure 4D) than in Bx2 (0.05%), whereas the proportions of CD163+ and CD163− macrophages and monocytes were higher in Bx2 (51.2%) than in Bx1 (18.9%), with the largest increase in CD163+ macrophages (Figure 4D). CD163 positivity is associated with differentiation of myelomonocytic cells toward an alternatively activated or “M2”-type state, which is considered to be pro-tumorigenic within solid tumors.30Barros M.H. Hauck F. Dreyer J.H. Kempkes B. Niedobitek G. Macrophage polarisation: an immunohistochemical approach for identifying M1 and M2 macrophages.PLoS One. 2013; 8: e80908.e8Crossref Scopus (320) Google Scholar,31Skytthe M.K. Graversen J.H. Moestrup S.K. Targeting of CD163(+) macrophages in inflammatory and Malignant diseases.Int. J. Mol. Sci. 2020; 21: 5497Crossref Scopus (24) Google Scholar CD163 expression on monocytes and macrophages is induced by interleukin-10 (IL-10) and glucocorticoids and repressed by lipopolysaccharides, tumor necrosis factor alpha (TNF-α), and interferon γ (IFNγ) and is concordant with upregulation of IL-containing GSVA gene sets in Bx2 (Figure 3C; Table S2).32Buechler C. Eisinger K. Krautbauer S. Diagnostic and prognostic potential of the macrophage specific receptor CD163 in inflammatory diseases.Inflamm. Allergy Drug Targets. 2013; 12: 391-402Crossref PubMed Scopus (47) Google Scholar The dominance of macrophages and monocytes and relative lack of T cells in the PT, Bx1, and Bx2 was in stark contrast to Bx4, which had many more T cells than macrophages and monocytes (PT: 65.8% macrophages/monocytes, 10.9% T cells; Bx1: 48.3%, 20.6%; Bx2: 82.4%, 5.3%; Bx4: 6.1%, 33.1%; Figures 4C and 4E, orange). Analyses of T cell subsets and functionality showed that only a small fraction of CD3+CD4+ and CD3+CD8+ T cells in the PT, Bx1, or Bx4 expressed the programmed cell death-1 (PD-1) protein, which is typically expressed on activated T cells following T cell priming or persistent antigen exposure (PT: 1.5% CD4+ T cells, 4.1% CD8+ T cells; Bx1: 1.3%, 1.6%; Bx4: 0%, 0.7%; compared with Bx2: 10.1%, 30.5%; Bx3: 11.3%, 6.3%; Figure 4F).33Pauken K.E. Wherry E.J. Overcoming T cell exhaustion in infection and cancer.Trends Immunol. 2015; 36: 265-276Abstract Full Text Full Text PDF PubMed Scopus (556) Google Scholar However, T cell status was markedly altered in Bx2 (Figure 4C; Table S2). Notably, although T cells were least abundant in Bx2 compared with Bx1 and Bx4, the largest fraction of CD3+CD4+ and CD3+CD8+ T cells expressing PD-1 was observed in Bx2 (Bx2: 10.1% CD4+ T cells, 30.5% CD8+ T cells; compared with Bx1: 1.3%, 1.6%; Bx3: 11.3%, 6.3%; Bx4: 0%, 0.7%; Figure 4F), coincident with relatively reduced FoxP3+CD4+ regulatory T (Treg) cells (Bx1: 4.0%, Bx2: 1.1%, Bx3: 9.3%, Bx4: 0.5%) and expanded Th17 CD4+ T cells (Bx1: 2.5%, Bx2: 26.1%, Bx3: 0%, Bx4: 0%; Figures 4G and 4H). Analyses of PD-1

The Impact of the hAPP695SW Transgene and Associated Amyloid-β Accumulation on Murine Hippocampal Biochemical Pathways

Abstract: Background: Alzheimer’s disease (AD) is a neurodegenerative disease characterized by the accumulation of amyloid-β (Aβ) peptide in the brain. Objective: To gain a better insight into alterations in major biochemical pathways underlying AD. Methods: We compared metabolomic profiles of hippocampal tissue of 20-month-old female Tg2576 mice expressing the familial AD-associated hAPP695SW transgene with their 20-month-old wild type female littermates. Results: The hAPP695SW transgene causes overproduction and accumulation of Aβ in the brain. Out of 180 annotated metabolites, 54 metabolites differed (30 higher and 24 lower in Tg2576 versus wild-type hippocampal tissue) and were linked to the amino acid, nucleic acid, glycerophospholipid, ceramide, and fatty acid metabolism. Our results point to 1) heightened metabolic activity as indicated by higher levels of urea, enhanced fatty acid β-oxidation, and lower fatty acid levels; 2) enhanced redox regulation; and 3) an imbalance of neuro-excitatory and neuro-inhibitory metabolites in hippocampal tissue of aged hAPP695SW transgenic mice. Conclusion: Taken together, our results suggest that dysregulation of multiple metabolic pathways associated with a concomitant shift to an excitatory-inhibitory imbalance are contributing mechanisms of AD-related pathology in the Tg2576 mouse.

The Eczema Area and Severity Index—A Practical Guide

Abstract: Atopic dermatitis is a chronic inflammatory skin condition that affects approximately 18 million people in the United States. Assessing the extent and severity of atopic dermatitis is critical for determining baseline disease burden and treatment effectiveness for both investigators and clinicians. Considerable efforts over the past several decades have been made in developing a highly validated instrument called the Eczema Area and Severity Index (EASI). Although several guides exist for the EASI, questions continue to arise regarding its use and interpretation. This review was developed to serve as the definitive guide for the EASI and to address commonly asked questions.

Graft Augmentation of Repairable Rotator Cuff Tears: An Algorithmic Approach Based on Healing Rates

Abstract: We provide our algorithm for tissue augmentation of rotator cuff repairs based on the current available evidence regarding rotator cuff healing. A variety of factors are associated with healing following rotator cuff repair. Increasing tear size and retraction as well as severe fatty degeneration have been associated with worsening rates of tendon healing. Given the correlation between tendon healing and postoperative outcomes, it is important to identify patients at high risk for failure and to modify their treatment accordingly to minimize the risk of early biomechanical failure and maximize the potential for structural healing. One approach that may be used to improve healing is tissue augmentation. Tissue augmentation is the use of tissue patches and scaffolds to provide rotator cuff reinforcement. Surgical management for rotator cuff tears (RCTs) continues to be a challenging task in orthopaedic surgery today. Appropriate treatment measures require an in depth understanding and consideration of the patient's prognostic factors such as age, fatty infiltration of the rotator cuff muscles, bone mineral density, rotator cuff retraction, anteroposterior tear size, work activity, and degenerative changes of the joint. Using these factors within the Rotator Cuff Healing Index, we can determine a patient's surgical treatment that will yield the maximum healing rate. For nonarthritic RCTs, joint-preserving strategies should be first-line treatment options. For young, active patients with a reparable RCT and minimal fatty infiltration, a complete repair can be effective. For young patients with irreparable RCTs, superior capsular reconstructions, and tendon transfers are viable options. For elderly patients with low work activity, an irreparable RCT and significant fatty infiltration, a partial repair with or without graft augmentation can be attempted if minimal to no arthritic changes are seen. LEVEL OF EVIDENCE: Level V, expert opinion.

Characterization of a Dynamic Y₂Ir₂O₇ Catalyst during the Oxygen Evolution Reaction in Acid

Abstract: Reducing precious metal content and improving the efficiency of proton exchange membrane water electrolyzers is critical for producing renewable hydrogen cost-effectively. Mixed metal iridium oxide catalysts (AIrxOy, A = nonprecious metal) have demonstrated superior oxygen evolution reaction (OER) activity relative to IrO2 catalysts while utilizing less Ir. However, improved stability is required if these materials are to be implemented commercially. In this work, we use a combination of ex situ and in situ characterization techniques to study physical and electronic properties of Y2Ir2O7 as it evolves during OER in acidic electrolyte. We identify and quantify dissolution of Y and Ir, finding that this material exhibits similar stability to other reported mixed metal Ir oxides (104–105 molO2 evolved/molIr dissolved) and appears to become more stable over time. We find that the catalyst surface becomes enriched with Ir after electrochemical testing. We further monitored the Ir oxidation state in situ using high-energy resolution fluorescence detected X-ray absorption spectroscopy. Our results suggest that the Ir oxidation state is dynamic: an IrOx surface forms that is more oxidized than the bulk pyrochlore material but subsequently dissolves. Such detailed characterization of material properties can be used to develop design principles for improving catalyst stability.

Central sympathetic network for thermoregulatory responses to psychological stress

Abstract: In mammals, many types of psychological stressors elicit a variety of sympathoexcitatory responses paralleling the classic fight-or-flight response to a threat to survival, including increased body temperature via brown adipose tissue thermogenesis and cutaneous vasoconstriction, and increased skeletal muscle blood flow via tachycardia and visceral vasoconstriction. Although these responses are usually supportive for stress coping, aberrant sympathetic responses to stress can lead to clinical issues in psychosomatic medicine. Sympathetic stress responses are mediated mostly by sympathetic premotor drives from the rostral medullary raphe region (rMR) and partly by those from the rostral ventrolateral medulla (RVLM). Hypothalamomedullary descending pathways from the dorsomedial hypothalamus (DMH) to the rMR and RVLM mediate important, stress-driven sympathoexcitatory transmission to the premotor neurons to drive the thermal and cardiovascular responses. The DMH also likely sends an excitatory input to the paraventricular hypothalamic nucleus to stimulate stress hormone release. Neurons in the DMH receive a stress-related excitation from the dorsal peduncular cortex and dorsal tenia tecta (DP/DTT) in the ventromedial prefrontal cortex. By connecting the corticolimbic emotion circuit to the central sympathetic and somatic motor systems, the DP/DTT → DMH pathway plays as the primary mediator of the psychosomatic signaling that drives a variety of sympathetic and behavioral stress responses. These brain regions together with other stress-related regions constitute a central neural network for physiological stress responses. This network model is relevant to understanding the central mechanisms by which stress and emotions affect autonomic regulations of homeostasis and to developing new therapeutic strategies for various stress-related disorders.

COVID-19 Vaccine Uptake Among People Living with HIV

Abstract: People living with HIV (PLWH) are at greater risk for severe COVID-19 and are a priority population for COVID-19 vaccination. As of June 15, 2021, 61.6% of PLWH in Oregon received ≥ 1 COVID-19 vaccine dose. Younger PLWH, Hispanic/Latinx PLWH and PLWH who inject drugs or reside in rural and frontier areas had low vaccine uptake while PLWH who were engaged in care, enrolled in the AIDS Drug Assistance Program, and vaccinated against influenza had high vaccine uptake. Greater advocacy, education, and care navigation are required to increase COVID-19 vaccine access and uptake among PLWH.

Effectiveness of COVID-19 treatment with nirmatrelvir-ritonavir or molnupiravir among U.S. Veterans: target trial emulation studies with one-month and six-month outcomes

Abstract: ABSTRACT Background Information about the effectiveness of oral antivirals in preventing short- and long-term COVID-19-related outcomes during the Omicron surge is limited. We sought to determine the effectiveness of nirmatrelvir-ritonavir and molnupiravir for the outpatient treatment of COVID-19. Methods We conducted three retrospective target trial emulation studies comparing matched patient cohorts who received nirmatrelvir-ritonavir versus no treatment, molnupiravir versus no treatment, and nirmatrelvir-ritonavir versus molnupiravir in the Veterans Health Administration (VHA). Participants were Veterans in VHA care at risk for severe COVID-19 who tested positive for SARS-CoV-2 in the outpatient setting during January and February 2022. Primary outcomes included all-cause 30-day hospitalization or death and 31-180-day incidence of acute or long-term care admission, death, or post-COVID-19 conditions. For 30-day outcomes, we calculated unadjusted risk rates, risk differences, and risk ratios. For 31-180-day outcomes, we used unadjusted time-to-event analyses. Results Participants were 90% male with median age 67 years and 26% unvaccinated. Compared to matched untreated controls, nirmatrelvir-ritonavir-treated participants (N=1,587) had a lower 30-day risk of hospitalization (27.10/1000 versus 41.06/1000, risk difference [RD] - 13.97, 95% CI -23.85 to -4.09) and death (3.15/1000 versus 14.86/1000, RD -11.71, 95% CI - 16.07 to -7.35). Among persons who were alive at day 31, further significant reductions in 31-180-day incidence of hospitalization (sub-hazard ratio 1.07, 95% CI 0.83 to 1.37) or death (hazard ratio 0.61, 95% CI 0.35 to 1.08) were not observed. Molnupiravir-treated participants aged ≥65 years (n=543) had a lower combined 30-day risk of hospitalization or death (55.25/1000 versus 82.35/1000, RD -27.10, 95% CI -50.63 to -3.58). A statistically significant difference in 30-day or 31-180-day risk of hospitalization or death was not observed between matched nirmatrelvir- or molnupiravir-treated participants. Incidence of most post-COVID conditions was similar across comparison groups. Conclusions Nirmatrelvir-ritonavir was highly effective in preventing 30-day hospitalization and death. Short-term benefit from molnupiravir was observed in older groups. Significant reductions in adverse outcomes from 31-180 days were not observed with either antiviral.

Quantitative Spatial Profiling of Immune Populations in Pancreatic Ductal Adenocarcinoma Reveals Tumor Microenvironment Heterogeneity and Prognostic Biomarkers

Abstract: Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive disease with poor 5-year survival rates, necessitating identification of novel therapeutic targets. Elucidating the biology of the tumor immune microenvironment (TiME) can provide vital insights into mechanisms of tumor progression. In this study, we developed a quantitative image processing platform to analyze sequential multiplexed IHC data from archival PDAC tissue resection specimens. A 27-plex marker panel was employed to simultaneously phenotype cell populations and their functional states, followed by a computational workflow to interrogate the immune contextures of the TiME in search of potential biomarkers. The PDAC TiME reflected a low-immunogenic ecosystem with both high intratumoral and intertumoral heterogeneity. Spatial analysis revealed that the relative distance between IL10+ myelomonocytes, PD-1+ CD4+ T cells, and granzyme B+ CD8+ T cells correlated significantly with survival, from which a spatial proximity signature termed imRS was derived that correlated with PDAC patient survival. Furthermore, spatial enrichment of CD8+ T cells in lymphoid aggregates was also linked to improved survival. Altogether, these findings indicate that the PDAC TiME, generally considered immuno-dormant or immunosuppressive, is a spatially nuanced ecosystem orchestrated by ordered immune hierarchies. This new understanding of spatial complexity may guide novel treatment strategies for PDAC.Quantitative image analysis of PDAC specimens reveals intertumoral and intratumoral heterogeneity of immune populations and identifies spatial immune architectures that are significantly associated with disease prognosis.

Circulating Cells with Macrophage-like Characteristics in Cancer: The Importance of Circulating Neoplastic-Immune Hybrid Cells in Cancer

Abstract: Cancer remains a significant cause of mortality in developed countries, due in part to difficulties in early detection, understanding disease biology, and assessing treatment response. If effectively harnessed, circulating biomarkers promise to fulfill these needs through non-invasive “liquid” biopsy. While tumors disseminate genetic material and cellular debris into circulation, identifying clinically relevant information from these analytes has proven difficult. In contrast, cell-based circulating biomarkers have multiple advantages, including a source for tumor DNA and protein, and as a cellular reflection of the evolving tumor. While circulating tumor cells (CTCs) have dominated the circulating cell biomarker field, their clinical utility beyond that of prognostication has remained elusive, due to their rarity. Recently, two novel populations of circulating tumor-immune hybrid cells in cancer have been characterized: cancer-associated macrophage-like cells (CAMLs) and circulating hybrid cells (CHCs). CAMLs are macrophage-like cells containing phagocytosed tumor material, while CHCs can result from cell fusion between cancer and immune cells and play a role in the metastatic cascade. Both are detected in higher numbers than CTCs in peripheral blood and demonstrate utility in prognostication and assessing treatment response. Additionally, both cell populations are heterogeneous in their genetic, transcriptomic, and proteomic signatures, and thus have the potential to inform on heterogeneity within tumors. Herein, we review the advances in this exciting field.

A Cancer Cell–Intrinsic GOT2–PPARδ Axis Suppresses Antitumor Immunity

Abstract: Abstract Despite significant recent advances in precision medicine, pancreatic ductal adenocarcinoma (PDAC) remains near uniformly lethal. Although immune-modulatory therapies hold promise to meaningfully improve outcomes for patients with PDAC, the development of such therapies requires an improved understanding of the immune evasion mechanisms that characterize the PDAC microenvironment. Here, we show that cancer cell–intrinsic glutamic-oxaloacetic transaminase 2 (GOT2) shapes the immune microenvironment to suppress antitumor immunity. Mechanistically, we find that GOT2 functions beyond its established role in the malate–aspartate shuttle and promotes the transcriptional activity of nuclear receptor peroxisome proliferator–activated receptor delta (PPARδ), facilitated by direct fatty acid binding. Although GOT2 is dispensable for cancer cell proliferation in vivo, the GOT2–PPARδ axis promotes spatial restriction of both CD4+ and CD8+ T cells from the tumor microenvironment. Our results demonstrate a noncanonical function for an established mitochondrial enzyme in transcriptional regulation of immune evasion, which may be exploitable to promote a productive antitumor immune response. Significance: Prior studies demonstrate the important moonlighting functions of metabolic enzymes in cancer. We find that the mitochondrial transaminase GOT2 binds directly to fatty acid ligands that regulate the nuclear receptor PPARδ, and this functional interaction critically regulates the immune microenvironment of pancreatic cancer to promote tumor progression. See related commentary by Nwosu and di Magliano, p. 2237.. This article is highlighted in the In This Issue feature, p. 2221

Long-term outcomes of osilodrostat in Cushing's disease: LINC 3 study extension

Abstract: Abstract Objective To investigate the long-term efficacy and tolerability of osilodrostat, a potent oral 11β-hydroxylase inhibitor, for treating Cushing's disease (CD). Design/methods A total of 137 adults with CD and mean 24-h urinary free cortisol (mUFC) > 1.5 × upper limit of normal (ULN) received osilodrostat (starting dose 2 mg bid; maximum 30 mg bid) during the prospective, Phase III, 48-week LINC 3 (NCT02180217) core study. Patients benefiting from osilodrostat at week 48 could enter the optional extension (ending when all patients had received ≥ 72 weeks of treatment or discontinued). Efficacy and safety were assessed for all enrolled patients from the core study baseline. Results Median osilodrostat exposure from the core study baseline to study end was 130 weeks (range 1–245) and median average dose was 7.4 mg/day (range 0.8–46.6). The reduction in mean mUFC achieved during the core was maintained during the extension and remained ≤ ULN. Of 106 patients, 86 (81%) patients who entered the extension had mUFC ≤ ULN at week 72. Improvements in cardiovascular/metabolic-related parameters, physical manifestations of hypercortisolism (fat pads, central obesity, rubor, striae, and hirsutism in females), and quality of life in the core study were also maintained or improved further during the extension. No new safety signals were reported; 15/137 (10.9%) and 12/106 (11.3%) patients discontinued for adverse events during the core and extension, respectively. Mean testosterone in females decreased towards baseline levels during the extension. Conclusions Data from this large, multicentre trial show that long-term treatment with osilodrostat sustains cortisol normalisation alongside clinical benefits in most patients with CD and is well tolerated.

Screening for Glaucoma in Adults

Abstract: Two 2013 systematic reviews to inform the US Preventive Services Task Force (USPSTF) found insufficient evidence to assess benefits and harms of screening for primary open-angle glaucoma (OAG) in adults.To update the 2013 reviews on screening for glaucoma, to inform the USPSTF.Ovid MEDLINE, the Cochrane Central Register of Controlled Trials, and the Cochrane Database of Systematic Reviews (to February 2021); surveillance through January 21, 2022.Randomized clinical trials (RCTs) of screening, referral, and treatment; and studies of screening test diagnostic accuracy.One investigator abstracted data and a second checked accuracy. Two investigators independently assessed study quality.Eighty-three studies (N = 75 887) were included (30 trials and 53 diagnostic accuracy studies). One RCT (n = 616) found screening of frail elderly persons associated with no difference in vision outcomes vs no screening but with significantly greater falls risk (relative risk [RR], 1.31 [95% CI, 1.13-1.50]). No study evaluated referral to an eye health professional. For glaucoma diagnosis, spectral domain optical coherence tomography (providing high-resolution cross-sectional imaging; 15 studies, n = 4242) was associated with sensitivity of 0.79 (95% CI, 0.75-0.83) and specificity of 0.92 (95% CI, 0.87-0.96) and the Humphrey Visual Field Analyzer (for perimetry, or measurement of visual fields; 6 studies, n = 11 244) with sensitivity of 0.87 (95% CI, 0.69-0.95) and specificity 0.82 (95% CI, 0.66-0.92); tonometry (for measurement of intraocular pressure; 13 studies, n = 32 892) had low sensitivity (0.48 [95% CI, 0.31-0.66]). Medical therapy for ocular hypertension and untreated glaucoma was significantly associated with decreased intraocular pressure and decreased likelihood of glaucoma progression (7 trials, n = 3771; RR, 0.68 [95% CI, 0.49-0.96]; absolute risk difference -4.2%) vs placebo, but 1 trial (n = 461) found no differences in visual acuity, quality of life, or function. Selective laser trabeculoplasty and medical therapy had similar outcomes (4 trials, n = 957).This review found limited direct evidence on glaucoma screening, showing no association with benefits. Screening tests can identify persons with glaucoma and treatment was associated with a lower risk of glaucoma progression, but evidence of improvement in visual outcomes, quality of life, and function remains lacking.

Two-Year Retrospective Patient-Reported Outcomes Following Superior Capsular Reconstruction

Abstract: The purpose of this study was to evaluate the short-term patient-reported outcomes of superior capsular reconstruction (SCR) and identify factors contributing to the success or failure of the procedure at 2 years.A retrospective review was performed on data prospectively collected from the Surgical Outcomes System database. Patient-reported outcomes (PROMs) including American Shoulder and Elbow Surgeons (ASES) score, Single Assessment Numeric Evaluation (SANE) score, visual analog scale for pain, and Veterans RAND 12-Item Health Survey (VR-12) were evaluated at a minimum of 2 years postoperatively and reported using a minimal clinically important difference (MCID) and the percent of maximal possible improvement (MPI). In addition, preoperative and intraoperative variables were evaluated in patients with and without a postoperative improvement in ASES and SANE scores meeting the threshold of MCID.Two-year follow-up data were available for 350 patients. Statistically significant improvements were noted in all PROMs at 2-year follow-up. In total, 240 patients (68.8%) achieved an MCID improvement of >17.5 in ASES score, and 185 patients (52.9%) achieved an MCID of >29.8 improvement in the SANE score. Primary SCRs were associated with a higher MPI in the ASES score (60.1% ± 39.8% vs 40.4% ± 47.9%; P = .025) and VR-12 physical score (14.0% ± 13.8% vs 8.0% ± 14.7%; P = .028) compared to revision repairs. Only diabetes was identified as a predictor of SANE score improvement (64.5% vs 62.2%; P = .041).SCR is associated with improvement in patient-reported outcomes at short-term follow-up, with 53% to 69% of patients achieving an improvement considered to meet the MCID. Greater improvement is expected when SCR is performed as a primary procedure rather than as a revision procedure for failed rotator cuff repair.Level III, retrospective comparative study.

A framework for multiplex imaging optimization and reproducible analysis

Abstract: Multiplex imaging technologies are increasingly used for single-cell phenotyping and spatial characterization of tissues; however, transparent methods are needed for comparing the performance of platforms, protocols and analytical pipelines. We developed a python software, mplexable, for reproducible image processing and utilize Jupyter notebooks to share our optimization of signal removal, antibody specificity, background correction and batch normalization of the multiplex imaging with a focus on cyclic immunofluorescence (CyCIF). Our work both improves the CyCIF methodology and provides a framework for multiplexed image analytics that can be easily shared and reproduced.

Tofacitinib for the treatment of active ankylosing spondylitis in adults

Abstract: Ankylosing spondylitis, also known as radiographic axial spondyloarthritis, is a complex, immune-mediated inflammatory disorder most commonly involving the spine including the sacroiliac joints.Complex pathogenesis of axial spondyloarthritis involving genetic, environmental, and both innate and adaptive immune systems. Treatment options for ankylosing spondylitis. Pharmacologic properties, efficacy, and safety of tofacitinib, a JAK inhibitor. Data regarding efficacy of approved JAK inhibitors in the treatment of ankylosing spondylitis, including tofacitinib, upadacitinib, and filgotinib.Current treatment options of ankylosing spondylitis include NSAIDs, TNFi, and IL-17i. JAK inhibitors present a new class of therapy that has shown efficacy in the treatment of active ankylosing spondylitis in adults. While it has not been directly compared to alternative therapies, tofacitinib has been shown to be effective in both phase II and phase III trials for the treatment of ankylosing spondylitis. While these trials did not show any significant difference from placebo in terms of safety, the ORAL Surveillance study showed tofacitinib to be inferior to TNFi when comparing adverse events. Thus, tofacitinib presents a viable treatment option for the management of AS, however shared decision-making regarding risks and benefits will be important.

Understanding Drug Sensitivity and Tackling Resistance in Cancer

Abstract: Decades of research into the molecular mechanisms of cancer and the development of novel therapeutics have yielded a number of remarkable successes. However, our ability to broadly assign effective, rationally targeted therapies in a personalized manner remains elusive for many patients, and drug resistance persists as a major problem. This is in part due to the well-documented heterogeneity of cancer, including the diversity of tumor cell lineages and cell states, the spectrum of somatic mutations, the complexity of microenvironments, and immune-suppressive features and immune repertoires, which collectively require numerous different therapeutic approaches. Here, we describe a framework to understand the types and biological causes of resistance, providing translational opportunities to tackle drug resistance by rational therapeutic strategies.

Assessing the risk of thromboembolism in cancer patients receiving immunotherapy

Abstract: Malignancy has long been implicated with hypercoagulability, leading to an increased rate of both venous and arterial thromboembolic events (VTE and ATE). Immunotherapy has established itself as a cornerstone of modern cancer therapy by promoting antitumor immune responses, though there have been some suggestions that immune-related adverse events could include increased rates of VTE and ATE. In this review, we examine the available evidence regarding the use of immune checkpoint inhibitors (ICIs) and thrombosis. First, we describe the potential mechanisms by which ICIs might lead to thrombophilia given the overlap between the immune system, coagulation cascade, and platelet adhesion and activation. In addition, while there are some preclinical data evaluating immunotherapy-associated ATEs in animal models, there is a paucity of evidence exploring potential mechanism of VTEs in ICIs. Second, we review the incidence of ATE and VTE in patients receiving ICIs in the published literature. Finally, we discuss current limitations in understanding, areas of conflicting evidence, and approaches to further investigation.

A Phase 1 Study of the Safety, Tolerability, and Pharmacokinetics of Single Ascending Doses of a First-in-Human Engineered Cationic Peptide, PLG0206, Intravenously Administered in Healthy Subjects

Abstract: In this first-in-human study, PLG0206, a novel engineered cationic antimicrobial peptide, was evaluated for safety, tolerability, and pharmacokinetics (PK) when intravenously (i.v.) administered as a single dose to healthy subjects. Six cohorts of 8 subjects each received escalating single i.v. infusions of PLG0206 at 0.05, 0.125, 0.25, 0.5, or 1 mg/kg dose or placebo over 1 to 4 h. Subjects were randomized to receive either PLG0206 (6 per cohort) or placebo (2 per cohort). Serial pharmacokinetic samples were taken prior to infusion and up to 48 h postinfusion. Safety and tolerability were assessed throughout the study. The demographic characteristics of subjects were comparable between those treated with PLG0206 and placebo and between dose groups. The incidence of treatment-emergent adverse events (TEAE) related to PLG0206 was low, and most events were mild in severity and were similar between the PLG0206 treatment and placebo groups. The most common adverse events reported for PLG0206 were infusion-related reactions, which were mitigated with increasing infusion time and volume. There were no severe adverse events (SAEs), life-threatening events, or deaths throughout the study. i.v. PLG0206 exhibited linear pharmacokinetics over the dose range of 0.05 to 1.0 mg/kg. The median terminal half-life (t1/2) ranged from 7.37 to 19.97 h. Following a single i.v. infusion to healthy subjects, PLG0206 was safe and well tolerated and exhibited linear PK at doses ranging from 0.05 to 1 mg/kg. These findings support the ongoing development of i.v. PLG0206 as an antimicrobial agent.