HLA-J, a Non-Pseudogene as a New Prognostic Marker for Therapy Response and Survival in Breast Cancer

The human leukocyte antigen (HLA) genes are cell-surface proteins, essential for immune cell interaction. HLA-G is known for their high immunosuppressive effect and its potential as predictive marker in breast cancer. However, nothing is known about the HLA-J and its immunosuppressive, prognostic and predictive features, as it is assumed to be a pseudogene by in silico sequence interpretation. HLA-J, ESR1, ERBB2, KRT5 and KRT20 mRNA expression were analysed in 29 fresh frozen breast cancer biopsies and their corresponding resectates obtained from patients treated with neoadjuvant chemotherapy (NACT). mRNA was analysed with gene specific TaqMan-based Primer/Probe sets and normalized to Calmodulin 2. All breast cancer samples did express HLA-J and frequently increased HLA-J mRNA levels after NACT. HLA-J mRNA was significantly associated with overexpression of the ESR1 mRNA status (Spearman ρ 0,5679; p = 0.0090) and KRT5 mRNA (Spearman ρ 0,6121; p = 0.0041) in breast cancer core biopsies and dominated in luminal B subtype. Kaplan Meier analysis revealed that an increase of HLA-J mRNA expression after NACT had worse progression free survival (p = 0,0096), indicating a counterreaction of tumor tissues presumably to prevent elimination by enhanced immune infiltration induced by NACT. This counterreaction is associated with worse prognosis. To our knowledge this is the first study identifying HLA-J as a new predictive marker in breast cancer being involved in immune evasion mechanisms.Humane Leukozyten-Antigene (HLA) sind Proteine auf der Zelloberfläche, die essenziell für die Immunzellinteraktion sind. HLA-G ist für seine hohe immunosuppressive Wirkung sowie als potenzieller prädikativer Marker für Brustkrebs bekannt. Dagegen ist kaum etwas über HLA-J und seine immunosuppressiven, prognostischen und prädiktiven Eigenschaften bekannt, da es basierend auf In-silico-Sequenzanalysen als „Pseudogen“ interpretiert wurde. Die Expression von HLA-J, ESR1, ERBB2, KRT5 und KRT20 mRNA wurde in 29 frisch gefrorenen Brustkrebsbiopsien analysiert und mit den klinisch-pathologischen Daten von Patientinnen, welche mit neoadjuvanter Chemotherapie behandelt wurden, verglichen. Die mRNA-Expression wurde mit genspezifischen TaqMan-basierten Primer/Probe-Sets analysiert und auf Calmodulin 2 normalisiert. Alle Gewebeproben von Patientinnen mit Brustkrebs exprimierten HLA-J, und der HLA-J-mRNA-Spiegel war nach NACT oft erhöht. In den Brustkrebsstanzbiopsien war die HLA-J-mRNA-Expression signifikant mit der Überexpression von ESR1-mRNA (Spearmans ρ 0,5679; p = 0,0090) und KRT5-mRNA (Spearmans ρ 0,6121; p = 0,0041) assoziiert und dominierte im Luminal-B-Subtyp. Die Kaplan-Meier-Analyse zeigte, dass ein Anstieg der HLA-J-mRNA-Expression nach NACT mit einem schlechteren progressionsfreien Überleben einhergeht (p = 0,0096), womöglich als Gegenreaktion des Tumorgewebes, um eine Eliminierung durch tumorinfiltrierende Lymphozyten, welche durch eine NACT induziert wurden, zu verhindern. Diese Gegenreaktion ist mit einer schlechteren Prognose assoziiert. Soweit uns bekannt, handelt es sich hierbei um die erste Studie, die HLA-J als neuen prädiktiven Marker im Brustkrebs identifiziert hat und möglicherweise zur Immunevasion beiträgt

Comparison of immunohistochemistry with PCR for assessment of ER, PR, and Ki-67 and prediction of pathological complete response in breast cancer

Background: Proliferation may predict response to neoadjuvant therapy of breast cancer and is commonly assessed by manual scoring of slides stained by immunohistochemistry (IHC) for Ki-67 similar to ER and PgR. This method carries significant intra- and inter-observer variability. Automatic scoring of Ki-67 with digital image analysis (qIHC) or assessment of MKI67 gene expression with RT-qPCR may improve diagnostic accuracy. Methods: Ki-67 IHC visual assessment was compared to the IHC nuclear tool (AperioTM) on core biopsies from a randomized neoadjuvant clinical trial. Expression of ESR1, PGR and MKI67 by RT-qPCR was performed on RNA extracted from the same formalin-fixed paraffin-embedded tissue. Concordance between the three methods (vIHC, qIHC and RT-qPCR) was assessed for all 3 markers. The potential of Ki-67 IHC and RT-qPCR to predict pathological complete response (pCR) was evaluated using ROC analysis and non-parametric Mann-Whitney Test. Results: Correlation between methods (qIHC versus RT-qPCR) was high for ER and PgR (spearman´s r = 0.82, p < 0.0001 and r = 0.86, p < 0.0001, respectively) resulting in high levels of concordance using predefined cut-offs. When comparing qIHC of ER and PgR with RT-qPCR of ESR1 and PGR the overall agreement was 96.6 and 91.4%, respectively, while overall agreement of visual IHC with RT-qPCR was slightly lower for ER/ESR1 and PR/PGR (91.2 and 92.9%, respectively). In contrast, only a moderate correlation was observed between qIHC and RT-qPCR continuous data for Ki-67/MKI67 (Spearman’s r = 0.50, p = 0.0001). Up to now no predictive cut-off for Ki-67 assessment by IHC has been established to predict response to neoadjuvant chemotherapy. Setting the desired sensitivity at 100%, specificity for the prediction of pCR (ypT0ypN0) was significantly higher for mRNA than for protein (68.9% vs. 22.2%). Moreover, the proliferation levels in patients achieving a pCR versus not differed significantly using MKI67 RNA expression (Mann-Whitney p = 0.002), but not with qIHC of Ki-67 (Mann-Whitney p = 0.097) or vIHC of Ki-67 (p = 0.131). Conclusion: Digital image analysis can successfully be implemented for assessing ER, PR and Ki-67. IHC for ER and PR reveals high concordance with RT-qPCR. However, RT-qPCR displays a broader dynamic range and higher sensitivity than IHC. Moreover, correlation between Ki-67 qIHC and RT-qPCR is only moderate and RT-qPCR with MammaTyper® outperforms qIHC in predicting pCR. Both methods yield improvements to error-prone manual scoring of Ki-67. However, RT-qPCR was significantly more specific

Clinical Predictors of Alcohol Withdrawal Delirium

Wetterling T, Kanitz R-D, Veltrup C, Driessen M. Clinical Predictors of Alcohol Withdrawal Delirium. Alcoholism: Clinical and Experimental Research. 1994;18(5):1100-1102.Up to now, clinical predictors for the course of the alcohol withdrawal syndrome, especially for the occurrence of a delirium, are lacking. Thus, this study was undertaken to examine whether clinical routine investigations at admission before the withdrawal syndrome can reveal factors indicating a higher risk for the development of a delirium. Our results showed that decreased serum electrolyte concentrations (i.e., chloride and potassium), elevated ALT, and γ-glutamyltransferase serum levels, as well as ataxia and polyneuropathy at the neurological examination, indicate a higher risk for the development of an alcohol withdrawal delirium

Kosten-Nutzen-Analyse klinisch-evaluierter Behandlungsprogramme

Driessen M, Veltrup C, Junghanns K, Przywara A, Dilling H. Kosten-Nutzen-Analyse klinisch-evaluierter Behandlungsprogramme. Der Nervenarzt. 1999;70(5):463-470.Kosten-Nutzen-Analysen neuer Therapieansätze ergänzen klinische Evaluationsstudien und erlauben eine bessere Gesamtabschätzung der Therapieeffizienz. Das Modell einer bereits klinisch evaluierten erweiterten Entzugsbehandlung Alkoholabhängiger (Entzug II) sollte unter Kosten-Nutzen-Gesichtspunkten überprüft werden. Es wurden 57 Patienten nach Entzug II und 37 nach konventioneller Entgiftung (Entzug I) untersucht. In einem retro- und prospektiven Ansatz wurden für 5 Jahre vor und nach Indextherapie Krankenversicherungsdaten zur Hospitalisierungshäufigkeit und -dauer, Arbeitsunfähigkeit und Krankengeldbezug erhoben. Entzug-II-Patienten wurden nach Indextherapie durchschnittlich seltener (3,5+4,4 vs. 7,3+11,3) und weniger lange (66+75 vs. 136+167) hospitalisiert und bezogen weniger lange Krankengeld (67+73 vs. 220+187) als Patienten nach Entzug I. Für den stationären Bereich ergaben sich bei Entzug-II-Patienten um ca. 50% geringere Gesamtbehandlungskosten (Indextherapie und Folgekosten). Insgesamt sprechen bei einer leichten klinischen Überlegenheit in den 12 Monaten nach Indextherapie (ca. 14% höhere Abstinenzrate) erheblich geringere Folgehospitalisierungen und Folgekosten für eine ausreichend hohe Effizienz der erweiterten Entzugstherapie Alkoholabhängiger

Psychiatric Comorbidity in Hospitalized Alcoholics after Detoxification Treatment

Driessen M, Arolt V, John U, Veltrup C, Dilling H. Psychiatric Comorbidity in Hospitalized Alcoholics after Detoxification Treatment. European Addiction Research. 1996;2(1):17-23.The aim of this study is to analyze psychiatric comorbidity of alcohol dependence with respect to prevalence rates, time of onset and indications for the course of alcoholism. 100 hospitalized alcohol-dependent patients were investigated. They were diagnosed according to the criteria of ICD-10 assessed by the Composite International Diagnostic Interview (CIDI). In 54% of the patients, at least one additional psychiatric disorder was diagnosed (lifetime). The current prevalence rates for all diagnoses were 49% (1 year) and 38% (2 weeks), respectively. In 71.2% the onset of comorbidity disorders preceded the onset of alcoholism by more than 1 year. In patients with secondary depressive disorders, alcohol-related problems had manifested significantly earlier than in primary depressive patients, but no differences were found in the maximum amount of drinking, the age of onset of excessive drinking and the amount of drinking during the year before the present study. ICD-10 comorbidity rates on the basis of the CIDI are in agreement with former DSM-III-R results based on the DIS. Psychiatric comorbidity seems to play a role in the early stages of alcoholism but not in the later course

MOTIVATIONAL INTERVENTION: AN INDIVIDUAL COUNSELLING VS A GROUP TREATMENT APPROACH FOR ALCOHOL-DEPENDENT IN-PATIENTS

John U, Veltrup C, Driessen M, Wetterling T, Dilling H. MOTIVATIONAL INTERVENTION: AN INDIVIDUAL COUNSELLING VS A GROUP TREATMENT APPROACH FOR ALCOHOL-DEPENDENT IN-PATIENTS. Alcohol and Alcoholism. 2003;38(3):263-269.Aims: The present study aimed to evaluate whether individual counselling for alcohol-dependent patients in three sessions is as effective as a 2-week group treatment programme as part of an in-patient stay in a psychiatric hospital which was to foster motivation to seek further help and to strengthen the motivation to stay sober. Of particular importance was the external validity of the results, i.e. a ‘normal’ intake load of in-patients in detoxification and a wide variety of motivation to stop drinking were to be investigated. Methods: Subjects eligible for the study were all patients with alcohol problems admitted to a psychiatric hospital, but without psychosis, as the main diagnosis, and with a maximum of 10 detoxification treatments in the past. A randomized-controlled trial was conducted with 161 alcohol-dependent in-patients who received three individual counselling sessions on their ward in addition to detoxification treatment and 161 in-patients who received 2 weeks of in-patient treatment and four out-patient group sessions in addition to detoxification. Both interventions followed the principles and strategies of motivational interviewing. Results: Six months after intervention, group-treatment patients showed a higher rate of participation in self-help groups; however, this difference had disappeared 12 months after treatment. The abstinence rate among the former patients did not differ between the two intervention groups. Conclusion: Group treatment may lead to a higher rate of participation in self-help groups, but does not increase the abstinence rate 6 months after treatment

Moisture behavior of weathered wood surfaces during cyclic wetting : measurements and modeling

The effects of weathering on the in-service moisture behavior of wood have received only limited attention so far, with much focus being on the effect of photodegradation on the hydrophobicity of the wood surface. The objective of the present study was to examine the effect of weathering surfaces on the overall moisture behavior of wood specimens exposed to short-term cyclic spraying, with special emphasis on the surface conditions involved. Specimens cut from eight different species including hardwoods and softwoods were weathered for 8 years and continuously monitored during a single-sided cyclic spraying together with a set of axially matched controls. After each spray cycle, the duration of surface wetness was evaluated by resistance moisture sensors as well as an optical approach (colorimetric) based on time-lapse images. The moisture content in the core was monitored simultaneously by use of resistance moisture sensors. The optical method correlated well with the electrical resistance measurements and provided a simple and practical measure of the areal distribution of the surface wetness. The results showed specimens with a weathered surface to sustain a wet surface for about twice the duration of their axially matched control. A considerable, albeit smaller, effect was also observed deeper in the core. By adapting the length of the wet period on the exposed boundary, the corresponding response at the core of the Norway spruce specimens was reproduced numerically