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Pulmonary hypertension (PH) is characterized by progressive obstruction of pulmonary arteries owing to inflammatory processes, cellular proliferation, and extracellular matrix deposition and vasoconstriction. As treatment options are limited, we studied gene transfer of an inducible nitric oxide synthase (iNOS) using adeno-associated virus (AAV) vectors specifically targeted at endothelial cells of pulmonary vessels in a murine model of PH. Adult mice were intravenously injected with AAV vectors expressing iNOS. Mice were subjected to hypoxia for 3 weeks and killed afterward. We found elevated levels of iNOS both in lung tissue and pulmonary endothelial cells in hypoxic controls that could be further increased by AAV-mediated iNOS gene transfer. This additional increase in iNOS was associated with decreased wall thickness of pulmonary vessels, less macrophage infiltration, and reduced molecular markers of fibrosis. Taken together, using a tissue-targeted approach, we show that AAV-mediated iNOS overexpression in endothelial cells of the pulmonary vasculature significantly decreases vascular remodeling in a murine model of PH, suggesting upregulation of iNOS as promising target for treatment of PH.

Supplemental Figures 1 to 6 from Cetuximab Resistance in Head and Neck Cancer Is Mediated by EGFR-K521 Polymorphism

Supplementary Figure 1. Inhibitory effect of Cetuximab determined by western blot analysis of 12 HNSCC cell line lysates. Supplementary Figure 2. Time course of EGFR phosphorylation in HNSCC cell line UT-SCC 8. Supplementary Figure 3. Correlation of total and membrane exposed EGFR in HNSCC cell lines. Supplementary Figure 4. IL-3 dependent Ba/F3 cells can be transformed to EGF dependence by stable transduction with EGFR and EGFRK521 genes. Supplementary Figure 5. Far-UV CD spectra of Fc-EGFR and Fc-EGFRK521. Supplementary Figure 6. ADCC induction in EGFR-transduced Ba/F3 cells by Cetuximab and CetuGEX{trade mark, serif}.

Supplementary Table 1 Patient characteristics including EGFRK521 allele frequency. Supplementary Table 2 Characteristics and sequencing data of squamous cell carcinoma cell lines.

Head and neck squamous cell carcinomas (HNSCC) exhibiting resistance to the EGFR-targeting drug cetuximab poses a challenge to their effective clinical management. Here, we report a specific mechanism of resistance in this setting based upon the presence of a single nucleotide polymorphism encoding EGFR-K521 (K-allele), which is expressed in >40% of HNSCC cases. Patients expressing the K-allele showed significantly shorter progression-free survival upon palliative treatment with cetuximab plus chemotherapy or radiation. In several EGFR-mediated cancer models, cetuximab failed to inhibit downstream signaling or to kill cells harboring a high K-allele frequency. Cetuximab affinity for EGFR-K521 was reduced slightly, but ligand-mediated EGFR activation was intact. We found a lack of glycan sialyation on EGFR-K521 that associated with reduced protein stability, suggesting a structural basis for reduced cetuximab efficacy. CetuGEX, an antibody with optimized Fc glycosylation targeting the same epitope as cetuximab, restored HNSCC sensitivity in a manner associated with antibody-dependent cellular cytotoxicity rather than EGFR pathway inhibition. Overall, our results highlight EGFR-K521 expression as a key mechanism of cetuximab resistance to evaluate prospectively as a predictive biomarker in HNSCC patients. Further, they offer a preclinical rationale for the use of ADCC-optimized antibodies to treat tumors harboring this EGFR isoform. Cancer Res; 77(5); 1188–99. ©2016 AACR.

Interleukin-2 (IL-2) and its receptor (IL-2R) play a major role in cellular immunity. The monoclonal antibodies basiliximab and daclizumab directed against the IL-2R subunit CD25 are widely used to prevent graft or host rejection after allogeneic tissue transplantation. Although these antibodies have been used for this purpose for many years, their common epitope within the CD25 protein is unknown. We screened a random phage display library to isolate peptides specifically binding to basiliximab. A striking amino acid sequence motif was enriched. This motif is homologous to the peptide ERIYHFV comprising amino acid positions 116 to 122 within the extracellular domain of CD25, suggesting that this is the basiliximab epitope. Basiliximab and daclizumab binding of selected phage was specific, as no binding was observed to isotype antibody controls. Phage binding could be inhibited by the cognate peptide. In cells expressing mutant CD25, binding of basiliximab was abolished when two or more amino acids of the suspected epitope were changed. In contrast, basiliximab binding remained unaffected in cells expressing CD25 versions with mutations outside this epitope. We therefore conclude that the (116)ERIYHFV(122) string within CD25 is the epitope recognized by basiliximab and daclizumab. This epitope overlaps with the interaction site of CD25 and IL-2, thus revealing the structural basis for the inhibition of IL-2R binding by this class of immunosuppressive antibodies. [Cancer Res 2007;67(8):3518–23]

Supplementary Table 1 and Figures 1-2 from Identification of Their Epitope Reveals the Structural Basis for the Mechanism of Action of the Immunosuppressive Antibodies Basiliximab and Daclizumab

Simple Summary In Molecular Tumor Boards (MTBs), clinicians and researchers discuss the biology of tumor samples from individual patients to find suitable therapies. MTBs have therefore become key elements of precision oncology programs. Patients living in urban areas with specialized medical centers can easily access MTBs. Dedicated efforts are necessary to also grant equal access for patients from rural areas. To address this challenge, the four German cancer centers in Würzburg, Erlangen, Regensburg and Augsburg collectively measured the regional efficacy of their MTBs. By jointly analyzing the residences of all MTB patients, we uncovered regional differences in our mostly rural catchment area. Mapping and further understanding these local differences—especially the underrepresented white spots—will help resolving inequalities in patient access to precision oncology. Our study represents a hands-on approach to assessing the regional efficacy of a precision oncology program. Moreover, this approach is transferable to other regions and clinical applications. Abstract (1) Background: molecular tumor boards (MTBs) are crucial instruments for discussing and allocating targeted therapies to suitable cancer patients based on genetic findings. Currently, limited evidence is available regarding the regional impact and the outreach component of MTBs; (2) Methods: we analyzed MTB patient data from four neighboring Bavarian tertiary care oncology centers in Würzburg, Erlangen, Regensburg, and Augsburg, together constituting the WERA Alliance. Absolute patient numbers and regional distribution across the WERA-wide catchment area were weighted with local population densities; (3) Results: the highest MTB patient numbers were found close to the four cancer centers. However, peaks in absolute patient numbers were also detected in more distant and rural areas. Moreover, weighting absolute numbers with local population density allowed for identifying so-called white spots—regions within our catchment that were relatively underrepresented in WERA MTBs; (4) Conclusions: investigating patient data from four neighboring cancer centers, we comprehensively assessed the regional impact of our MTBs. The results confirmed the success of existing collaborative structures with our regional partners. Additionally, our results help identifying potential white spots in providing precision oncology and help establishing a joint WERA-wide outreach strategy.

BackgroundMolecular Tumor Boards (MTBs) are crucial instruments for discussing and allocating targeted therapies to suitable cancer patients based on genetic findings. Currently, limited evidence is available regarding the regional impact and the outreach component of MTBs.MethodsWe analyzed MTB patient data from four neighboring Bavarian tertiary care oncology centers in Würzburg, Erlangen, Regensburg, and Augsburg, together constituting the WERA Alliance. Absolute patient numbers and regional distribution across the WERA-wide catchment area were weighted with local population densities.ResultsHighest MTB patient numbers were found close to the four cancer centers. However, peaks in absolute patient numbers were also detected in more distant and rural areas. Moreover, weighting absolute numbers with local population density allowed us to identify regions within our catchment area relatively underrepresented in WERA MTBs.ConclusionInvestigating patient data from four neighboring cancer centers, we comprehensively assessed the regional impact of our MTBs. The results confirmed the success of existing collaborative structures with our regional partners. Additionally, our results help identifying potential white spots in precision oncology and establishing a joint WERA-wide outreach strategy.

While many authors have described the adverse health effects of poor air quality and meteorological extremes, there remain inconsistencies on a regional scale as well as uncertainty about the single and joint effects of atmospheric predictors. In this context, we investigated the short-term impacts of weather and air quality on moderate extreme cancer-related mortality events for the urban area of Augsburg, Southern Germany, during the period 2000–2017. First, single effects were uncovered by applying a case-crossover routine. The overall impact was assessed by performing a Mann–Whitney U testing scheme. We then compared the results of this procedure to extreme noncancer-related mortality events. In a second step, we found periods with contemporaneous significant predictors and carried out an in-depth analysis of these joint-effect periods. We were interested in the atmospheric processes leading to the emergence of significant conditions. Hence, we applied the Principal Component Analysis to large-scale synoptic conditions during these periods. The results demonstrate a strong linkage between high-mortality events in cancer patients and significantly above-average levels of nitrogen dioxide (NO2) and particulate matter (PM2.5) during the late winter through spring period. These were mainly linked to northerly to easterly weak airflow under stable, high-pressure conditions. Especially in winter and spring, this can result in low temperatures and a ground-level increase and the accumulation of air pollution from heating and traffic as well as eastern lateral advection of polluted air. Additionally, above-average temperatures were shown to occur on the days before mortality events from mid-summer through fall, which was also caused by high-pressure conditions with weak wind flow and intense solar radiation. Our approach can be used to analyse medical data with epidemiological as well as climatological methods while providing a more vivid representation of the underlying atmospheric processes.

Chemotherapy is still the backbone of systemic treatment in the majority of cancers. However, immunotherapies, especially those based on checkpoint inhibition, are additional therapy options for many. For this, functional T cells are a mandatory requirement. The aim of this prospective study was to investigate the influence of chemotherapy on the cellular immune status of individual patients. Peripheral blood samples of 26 patients with solid malignancies undergoing chemotherapy were analyzed for lymphocyte populations and their subsets in a longitudinal approach. Chemotherapy decreased total B lymphocyte counts [median value (25-75 percentile): before chemotherapy 76/µl (39-160) vs. after chemotherapy 49/µl (24-106); p = 0.001]. Among B cells, specific subsets decreased particularly [naïve B cells (49/µl (21-111) vs. 25/µl (13-56); p = 0.001], memory B cells [3/µl (2-8) vs. 2/µl (1-4); p = 0.001], and class-switched B cells [11/µl (6-20) vs. 6/µl (3-12); p = 0.011]. In contrast, chemotherapy had no influence on the total numbers of CD4 + and CD8 + T lymphocytes or on their subsets (T helper cells 1, 2, and 17 as well as cytotoxic T cells in early, intermediate, late, terminal effector and exhausted status as well as both T-cell types with naïve, center memory, effector memory, activated, or regulatory phenotype). Furthermore, the count of natural killer (NK) lymphocytes showed no significant change before and after chemotherapy. In summary, this study shows a decrease of B lymphocytes during systemic chemotherapy, but no relevant effect on T lymphocytes, NK lymphocytes and their subsets. This could support the idea of an effective additive T-cell-dependent immunotherapy to chemotherapy.

Parallel zur gestiegenen Uberlebenswahrscheinlichkeit der Patienten mit malignen Tumorerkrankungen stieg in den letzten Jahren auch die Zahl der Patienten mit Metastasen — sehr haufig sind dies komplexe Knochenmetastasen zum Beispiel in der Wirbelsaule. Fur diese multidisziplinare therapeutische Herausforderung bietet das NOMS-Framework einen zuverlassigen Algorithmus.

The human Alzheimer’s disease (AD) brain accumulates angiogenic markers but paradoxically, the cerebral microvasculature is reduced around Aß plaques. Here we demonstrate that angiogenesis is started near Aß plaques in both AD mouse models and human AD samples. However, endothelial cells express the molecular signature of non-productive angiogenesis (NPA) and accumulate, around Aß plaques, a tip cell marker and IB4 reactive vascular anomalies with reduced NOTCH activity. Notably, NPA induction by endothelial loss of presenilin, whose mutations cause familial AD and which activity has been shown to decrease with age, produced a similar vascular phenotype in the absence of Aß pathology. We also show that Aß plaque-associated NPA locally disassembles blood vessels, leaving behind vascular scars, and that microglial phagocytosis contributes to the local loss of endothelial cells. These results define the role of NPA and microglia in local blood vessel disassembly and highlight the vascular component of presenilin loss of function in AD., A.E.R.-N. was the recipient of a JdlC-F fellowship from the Spanish Ministry of Economy, Industry, and Competitiveness (MINEICO) (FJCI-2015-23708), M.I.A.-V., N.L.-U., and C.O.-d.S.L. were the recipient of an FPU fellowship from Spanish Ministry of Education, Culture, and Sport (respectively, FPU15/02898, FPU14-02115, and AP2010‐1598), and R.M.-D. was the recipient of a “Sara Borrell” fellowship from ISCIII (CD09/0007). Work was supported by grants to A.P. by the Spanish MINEICO, ISCIII, and FEDER (SAF2012‐33816, SAF2015‐64111‐R, RTI2018-096629-B-100, SAF2017-90794-REDT, and PIE13/0004), by the regional Government of Andalusia (“Proyectos de Excelencia”, P12‐CTS‐2138 and P12‐CTS‐2232) co-funded by CEC and FEDER funds, and by the “Ayuda de Biomedicina 2018”, Fundación Domingo Martínez; J.Vitorica: Instituto de Salud Carlos III (ISCiii) of Spain, co-financed by FEDER funds from European Union (PI18/01556) by La Marató-TV3 Foundation grant 20141431; by CIBERNED (CB06/05/0094); and by Junta de Andalucia Consejería de Economía y Conocimiento through grant US-1262734; A.G.: Instituto de Salud Carlos III (ISCiii) of Spain, co-financed by FEDER funds from European Union, through grant PI18/01557; and by Junta de Andalucia Consejería de Economía y Conocimiento through grants UMA18-FEDERJA-211 and P18-RT-2233 co-financed by Programa Operativo FEDER 2014-2020.

Triple-negative breast cancer (TNBC) is an aggressive tumor with limited treatment options and poor prognosis. We applied the in vivo phage display technology to isolate peptides homing to the immunosuppressive cellular microenvironment of TNBC as a strategy for non-malignant target discovery. We identified a cyclic peptide (CSSTRESAC) that specifically binds to a vitamin D receptor, protein disulfide-isomerase A3 (PDIA3) expressed on the cell surface of tumor-associated macrophages (TAM), and targets breast cancer in syngeneic TNBC, non-TNBC xenograft, and transgenic mouse models. Systemic administration of CSSTRESAC to TNBC-bearing mice shifted the cytokine profile toward an antitumor immune response and delayed tumor growth. Moreover, CSSTRESAC enabled ligand-directed theranostic delivery to tumors and a mathematical model confirmed our experimental findings. Finally, in silico analysis showed PDIA3-expressing TAM in TNBC patients. This work uncovers a functional interplay between a cell surface vitamin D receptor in TAM and antitumor immune response that could be therapeutically exploited.

Ever declining autopsy rates have been a concern of pathologists as well as clinicians for decades. Notably, in the field of oncology, data on autopsies and discrepancies between clinical and autoptic diagnoses are particularly scarce. In this retrospective study, we show the effect of a simple catalog of measures consisting of a different approach to obtain consent for autopsy, structured conferencing, and systematic teaching of residents, as well as a close collaboration between clinicians and pathologists on the numbers of autopsies, especially of oncological patients. Additionally, postmortem examination protocols from the years 2015 until 2019 were analyzed, regarding rates of discrepancies between clinical and autoptic causes of death in this category of patients. Autopsy numbers could be significantly increased from a minimum in 2014 (60 autopsies) to a maximum in 2018 (142 autopsies) (p

Aim Right- and left-sided primary tumors of colorectal origin differ substantially in several aspects. Recent retrospective analyses show distinct efficacy of EGFR-epidermal growth factor receptor (EGFR)-directed therapies for left- and right-sided primary tumors. Current treatment guidelines have accommodated these findings such that for right-sided primary tumors, EGFR-directed therapy is no longer recommended. Instead, vascular endothelial growth factor (VEGF)-directed therapies are recommended frequently in first line, even in tumors with wild-type rat sarcoma (RAS) status. However, data supporting this recommendation are scarce. The purpose of this analysis was to investigate the efficacy of bevacizumab added to chemotherapy depending on the primary tumor localization in a retrospective setting. Patients and methods From the central clinical cancer registry of one of Germany's largest medical centers, data were analyzed for patients with metastatic colorectal cancer (mCRC) treated with either chemotherapy alone (CT) or bevacizumab-containing regimens (BEV/CT). Results Of 1,080 documented mCRC cases within the period of 2003 through 2016, 242 were treated with chemotherapy alone and 166 with bevacizumab-containing regimes in any line of therapy meeting the criterion above. In patients with left-sided primary tumor localization, a significant survival benefit was found when bevacizumab was added to chemotherapy. Patients with right-sided primaries, instead, did not derive any advantage when bevacizumab was added to chemotherapy. For the whole group of patients, this translated into a trend towards improved survival in bevacizumab-treated patients with mCRC. Conclusion Adding bevacizumab to chemotherapy in mCRC may be beneficial only in patients with left-sided primary tumor, while those with right-sided primary tumors may have no additional benefit from the addition of bevacizumab. This hypothesis-generating analysis should provide a basis for in-depth analysis of this issue in future prospective trials.

Objective: Incontinentia pigmenti (IP) is a genetic disease leading to severe neurological symptoms, such as epileptic seizures, but no specific treatment is available. IP is caused by pathogenic variants that inactivate the Nemo gene. Replacing Nemo through gene therapy might provide therapeutic benefits. Methods: In a mouse model of IP we administered a single intravenous dose of the AAV vector AAV-BR1-CAG-NEMO delivering the Nemo gene to the brain endothelium. Spontaneous epileptic seizures and the integrity of the BBB were monitored. Results: The endothelium-targeted gene therapy improved the integrity of the blood-brain barrier. In parallel, it reduced the incidence of seizures and delayed their occurrence. Neonate mice intravenously injected with the AAV-BR1-CAG-NEMO vector developed no hepatocellular carcinoma or other major adverse effects 11 months after vector injection, demonstrating that the vector has a favorable safety profile. Interpretation: The data show that the blood-brain barrier is a target of antiepileptic treatment and more specifically provide evidence for the therapeutic benefit of a brain endothelial-targeted gene therapy in IP. This article is protected by copyright. All rights reserved.

This case series describes autopsy findings in 10 patients with proven severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection who died at a university medical center in Germany.

Background In recent years, increasing evidence of second neoplasms associated with gastrointestinal stromal tumors (GIST) has been found. Numerous case reports, mostly retrospective studies and a few reviews, have been published. To our knowledge, however, no systematic review or meta-analysis of the existing data has been performed so far. Aim To prepare a compilation, as complete as possible, of all reported second tumor entities that have been described in association with GIST and to systematically analyze the published studies with regard to frequency, localization, and types of GIST-associated neoplasms. Methods The MEDLINE and EBSCO databases were searched for a combination of the keywords GIST/secondary, synchronous, coincident/tumor, neoplasm, and relevant publications were selected by two independent authors. Results Initially, 3042 publications were found. After deletion of duplicates, 1631 remained, and 130 papers were selected; 22 of these were original studies with a minimum of 20 patients, and 108 were case reports. In the 22 selected studies, comprising a total number of 12050 patients, an overall rate of GIST-associated neoplasias of 20% could be calculated. Most second neoplasias were found in the gastrointestinal tract (32%) and in the male and female urogenital tract (30%). The specific risk scores of GISTs associated with other tumors were significantly lower than those without associated neoplasias. Conclusion In this first systematic review, we could confirm previously reported findings of a more than coincidental association between GIST and other neoplasias. The question whether there is an underlying causal association will need further investigation. Our data suggest that even GIST with a very low risk of disease progression should prompt screening for second neoplasia and subsequent frequent controls or extended staging.

Objective. Resection remains the most important form of treatment for patients with intracranial malignancies. The advantages of surgical resection must be balanced with the risk of surgery. Intraoperative neuromonitoring (IOM) is utilized to improve safety and enlarge the resection extent. However, comparative data on the use if IOM is scarce. Methods. Retrospective analysis of patients following resection of motor eloquent lesions between June 2018 and June 2020 was performed. A comparative analysis of the extend of resection; neurological squeal were compared between resections with or without IOM. Results. 66 patients (31 female, 35 male, age median, SD 62+/− 11,2) were identified. IOM was performed in 34 (51%) cases. 46% were female with a median age of 62 (range 31–82). There were no differences in baseline patient“s characteristics between both groups. 14% and 3% had a temporary neurological deterioration in the IOM and non-IOM group (P = 0,198), respectively. 3% and 15% had a permanent neurological in the IOM and non-IOM group, respectively (P = 0.100). 65% and 35% of resections were total and subtotal in both the IOM and non-IOM group (P = 0.027), respectively. Conclusion. IOM is associated with lower rates of new postoperative neurological deterioration with the same rate resection quality following resection of motor eloquent lesions.

Head and neck squamous cell carcinomas (HNSCC) exhibiting resistance to the EGFR-targeting drug cetuximab poses a challenge to their effective clinical management. Here, we report a specific mechanism of resistance in this setting based upon the presence of a single nucleotide polymorphism encoding EGFR-K521 (K-allele), which is expressed in >40% of HNSCC cases. Patients expressing the K-allele showed significantly shorter progression-free survival upon palliative treatment with cetuximab plus chemotherapy or radiation. In several EGFR-mediated cancer models, cetuximab failed to inhibit downstream signaling or to kill cells harboring a high K-allele frequency. Cetuximab affinity for EGFR-K521 was reduced slightly, but ligand-mediated EGFR activation was intact. We found a lack of glycan sialyation on EGFR-K521 that associated with reduced protein stability, suggesting a structural basis for reduced cetuximab efficacy. CetuGEX, an antibody with optimized Fc glycosylation targeting the same epitope as cetuximab, restored HNSCC sensitivity in a manner associated with antibody-dependent cellular cytotoxicity rather than EGFR pathway inhibition. Overall, our results highlight EGFR-K521 expression as a key mechanism of cetuximab resistance to evaluate prospectively as a predictive biomarker in HNSCC patients. Further, they offer a preclinical rationale for the use of ADCC-optimized antibodies to treat tumors harboring this EGFR isoform. Cancer Res; 77(5); 1188–99. ©2016 AACR.

Ein 35-jahriger Patient mit periodischem Fieber bis 40 °C, Halsschmerzen, Myalgie und Zephalgien stellte sich zur Abklarung einer Panzytopenie vor. Die Umfeld- und jungere Reiseanamnese waren, abgesehen von einem Besuch in Italien vor einem Jahr und in Spanien vor mehreren Jahren, leer. Neben der Panzytopenie mit Agranulozytose und erhohten Infektwerten ohne wegweisenden serologischen Befund fand sich klinisch eine Lymphadenitis colli. In der Schnittbildgebung fanden sich eine zervikale Lymphadenopathie, eine Hepatosplenomegalie sowie eine gedeckte Sigmaperforation mit parakolischem Abszess und langstreckiger diffuser Wandverdickung des Kolons. Im Knochenmark (KM) fand sich eine reaktive, polyklonale Plasmazellvermehrung. Im Lymphknotenbiopsat ergab sich das Bild einer nekrotisierenden Lymphadenitis. Im Lymphknoten liesen sich mittels Polymerase-Kettenreaktion Leishmanien-DNA und im KM-Ausstrichpraparat Leishmanien lichtmikroskopisch nachweisen. Somit konnte die Diagnose einer viszeralen Leishmaniose (VL) gestellt werden. Es wurde eine Therapie mit liposomalem Amphotericin B begonnen. Fieberschube und Lymphadenopathie zeigten sich daraufhin regredient. Die VL ist eine pleiotrop, haufig schwer und unbehandelt todlich, verlaufende Erkrankung, die sich ahnlich einer hamatologischen Systemerkrankung prasentiert, gelegentlich ein fulminantes Makrophagenaktivierungssyndrom auslost und weltweit in seiner Haufigkeit zunimmt. Pathophysiologisch liegt ein komplexes Zusammenspiel zwischen Erreger und Immunsystem vor.

Arterial occlusive events (AOEs) such as cerebrovascular, cardiovascular and peripheral arterial events are known side effects of ponatinib, assumed due to the rapid development and increase of arteriosclerosis, while the definitive pathomechanisms therefore are still unclear. We present a case of clinically apparent large vessel vasculitis and discuss this phenomenon as a possible mechanism of AOEs beside arteriosclerosis. J Hematol. 2019;8(2):83-85 doi: https://doi.org/10.14740/jh519