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Die malignen Speicheldrusentumoren sind eine extrem heterogene Gruppe seltener Tumoren mit groser klinischer, pathologischer und molekularbiologischer Variabilitat. Die chirurgische Resektion stellt in den meisten Fallen die Therapie der Wahl dar. Bei lokal fortgeschrittenen oder aggressiven malignen Tumoren ist haufig eine adjuvante Radiotherapie indiziert, die das Risiko fur ein Lokalrezidiv reduziert. Kontrovers diskutiert wird das Ausmas der Parotidektomie bei Karzinomen der Glandula parotis, insbesondere die Behandlung des N. facialis. Einigkeit herrscht weitestgehend daruber, diesen Nerv, wann immer moglich, zu erhalten, sofern nicht eine praoperative Parese oder eine offensichtliche intraoperative Tumorinfiltration einem Nervenerhalt entgegenstehen. Resezierte Anteile des N. facialis sollten moglichst sofort rekonstruiert werden. Die Chirurgie der von den kleinen Speicheldrusen ausgehenden Karzinome entspricht weitgehend dem Vorgehen bei Kopf-Hals-Plattenepithelkarzinomen. Bei adenoidzystischen Karzinomen ist aufgrund des perineuralen Wachstums und des haufigen Bezugs zur Schadelbasis eine R0-Resektion oft nicht zu erreichen, was die besondere Bedeutung der adjuvanten Radiotherapie bei dieser Entitat unterstreicht. Die Indikation der elektiven „neck dissection“ im cN0-Fall wird ebenfalls uneinheitlich gestellt. Wahrend einige Autoren die generelle Lymphknotenausraumung bei allen Karzinomen fordern, lautet der uberwiegende Konsens, eine elektive „neck dissection“ bei fortgeschrittener Tumorgrose, High-grade-Karzinomen und erhohtem Sicherheitsbedurfnis durchzufuhren. Die zunehmende Erkenntnis genomischer Alterationen und tumorspezifischer Signalwege eroffnet neue vielversprechende Moglichkeiten zielgerichteter molekularer Therapien im fortgeschrittenen Tumorstadium.

Purpose Despite promising achievements in precision cancer medicine (PCM), participating patients are still faced with manifold uncertainties, especially regarding a potential treatment benefit of molecular diagnostics (MD). Hence, MD poses considerable challenges for patient information and communication. To meet these challenges, healthcare professionals need to gain deeper insight into patients’ subjective experiences. Therefore, this qualitative study examined information aspects of MD programs in cancer patients. Methods In two German Comprehensive Cancer Centers, 30 cancer patients undergoing MD participated in semi-structured interviews on information transfer and information needs regarding MD. Additionally, patients provided sociodemographic and medical data and indicated their subjective level of information (visual analogue scale, VAS, 0–10). Results On average patients had high levels of information (mean = 7, median = 8); nevertheless 20% (n = 6) showed an information level below 5 points. Qualitative analysis revealed that patients show limited understanding of the complex background of MD and have uncertainties regarding their personal benefit. Further, patients described unmet information needs. Existential threat in awaiting the results was experienced as burdensome. To withstand the strains of their situation, patients emphasized the importance of trusting their physician. Conclusion The challenges in PCM consist in providing unambiguous information, especially concerning treatment benefit, and providing guidance and support. Therefore, psycho-oncology needs to develop guidelines for adequate patient communication in order to help healthcare providers and cancer patients to handle these challenges in the developing field of PCM.

Background Diagnosis and treatment decisions in cancer increasingly depend on a detailed analysis of the mutational status of a patient’s genome. This analysis relies on previously published information regarding the association of variations to disease progression and possible interventions. Clinicians to a large degree use biomedical search engines to obtain such information; however, the vast majority of scientific publications focus on basic science and have no direct clinical impact. We develop the Variant-Information Search Tool (VIST), a search engine designed for the targeted search of clinically relevant publications given an oncological mutation profile. Results VIST indexes all PubMed abstracts and content from ClinicalTrials.gov. It applies advanced text mining to identify mentions of genes, variants and drugs and uses machine learning based scoring to judge the clinical relevance of indexed abstracts. Its functionality is available through a fast and intuitive web interface. We perform several evaluations, showing that VIST’s ranking is superior to that of PubMed or a pure vector space model with regard to the clinical relevance of a document’s content. Conclusion Different user groups search repositories of scientific publications with different intentions. This diversity is not adequately reflected in the standard search engines, often leading to poor performance in specialized settings. We develop a search engine for the specific case of finding documents that are clinically relevant in the course of cancer treatment. We believe that the architecture of our engine, heavily relying on machine learning algorithms, can also act as a blueprint for search engines in other, equally specific domains. VIST is freely available at https://vist.informatik.hu-berlin.de/ Electronic supplementary material The online version of this article (10.1186/s12859-019-2958-3) contains supplementary material, which is available to authorized users.

T-box transcription factors, T-box expressed in T cells (T-bet) encoded by Tbx21 and Eomesodermin (Eomes), drive the differentiation of effector/memory T cell lineages and NK cells. The aim of the study was to determine the prognostic influence of the expression of these transcription factors in peripheral blood (pB) in a cohort of 41 metastatic (m) RCC patients before receiving sorafenib treatment and to analyze their association with the immunophenotype in pB. In contrast to Tbx21, in the multivariate analysis including clinical features, Eomes mRNA expression was identified as an independent good prognostic factor for progression-free survival (PFS, p = 0.042) and overall survival (OS, p = 0.001) in addition to a favorable ECOG performance status (p = 0.01 and p = 0.008, respectively). Eomes expression correlated positively not only with expression of Tbx21 and TGFβ1 mRNA, but also with mRNA expression of the activation marker ICOS, and with in vivo activated HLA-DR(+) T cells. Eomes expression was negatively associated with TNFα-producing T cells. On protein level, Eomes was mainly expressed by CD56(+)CD3(-) NK cells in pB. In conclusion, we identified a higher Eomes mRNA expression as an independent good prognostic factor for OS and PFS in mRCC patients treated with sorafenib.

Purpose The purpose of the trial was to assess the efficacy and tolerability of Sym004, a novel 1:1 mixture of two chimeric monoclonal antibodies (992 and 1024) targeting non-overlapping epitopes of the anti-epidermal growth factor receptor (EGFR), in patients with squamous cell carcinoma of the head and neck (SCCHN). Methods Incurable, recurrent and/or metastatic SCCHN patients with acquired resistance to anti-EGFR monoclonal antibody-containing treatment received weekly infusions of 12 mg/kg Sym004 until disease progression or unacceptable toxicity. Results Among the 26 patients treated with Sym004, the proportion of patients alive without disease progression at 6 months was 12 % (95 % CI 139 %). The median duration of progression-free survival was 82 days (95 % CI 41140 days). Of 19 patients evaluable for response, eight showed a decrease in the sum of the largest diameter in their target lesions (median 11 %; range 727 %). The best overall response was stable disease in 13 patients (50 %). Paired biopsies showed a significant down-regulation of EGFR in both skin and tumors following exposure to Sym004. All patients had EGFR-related adverse events, including grade 3 skin toxicities and grade ≥3 hypomagnesemia reported in 13 (50 %) and 10 (38 %) of 26 patients, respectively. One event fulfilling the protocol-defined criteria for infusion-related reactions (grade 2) was reported. No anti-drug antibodies were detected. Conclusions The marked EGFR down-regulation shown in target tissues supports the proposed mechanism of action of Sym004. This trial revealed modest anti-tumor activity of Sym004 in extensively pretreated advanced SCCHN patients.

Hintergrund Ein Schub in der Entwicklung neuer Ansatze zur medikamentosen Therapie der Plattenepithelkarzinome der Kopf-Hals-Region (SCCHN) hat sich in den letzten Jahren auf der Basis eines erheblich tieferen Verstandnisses der Tumorbiologie ergeben, wahrend die medikamentose Behandlung der anderen histologischen Subtypen nach wie vor schwierig bleibt. Dieser Beitrag fasst neuer medikamentose Ansatze, die in klinischer Evaluation sind, zusammen.

T cell based immunotherapy has been investigated in a variety of malignancies and analyses have been mostly founded on in vitro data with tumor cell monolayers. However, three-dimensional (3D) culture models might mimic more closely the 'in vivo' conditions than 2D monolayers. Therefore, we analyzed the expression of tumor-associated antigens (TAA) and of molecules involved in antigen processing and presentation (APM) in tumor spheres, which served as an in vitro model for micrometastasis which might be enriched in tumor propagating cancer stem cells. For enrichment of sphere cells 12 human solid tumor cell lines were cultured in serum-free medium. Expression of a variety of TAA and APM were analyzed by RT-PCR and/or flow cytometry and compared to expression in corresponding adherent bulk cells grown in regular growth medium. Compared to adherent cells, spheres showed equal or higher mRNA expression levels of LMP2, LMP7 and MECL-1, of TAP1 and TAP2 transporters and, surprisingly, also of TAA including differentiation antigens. However, downregulation or loss of HLA-I and HLA-II molecules in spheres was observed in 8 of 10 and 1 of 2 cell lines, respectively, and was unresponsive to stimulation with IFN-γ. Although tumor spheres express TAA and molecules of intracellular antigen processing, they are defective in antigen presentation due to downregulation of HLA surface expression which may lead to immune evasion.

Clinically effective T-cell responses can be elicited by single peptide vaccination with Wilms' tumor 1 (WT1) epitope 126-134 in patients with acute myeloid leukemia (AML). We recently showed that a predominant T-cell receptor (TCR) β chain was associated with vaccine-induced complete remission in an AML patient (patient 1). In this study, we address the question of whether this predominant clone or the accompanying Vβ11 restriction could be found in other AML patients vaccinated with the same WT1 peptide.For assessment of Vβ usage, cytotoxic T lymphocytes (CTLs) from four vaccinated patients were divided into specific and non-specific by epitope-specific enrichment. Vβ families were quantified in both fractions using reverse transcribed quantitative PCR. Vβ11-positive 'complementary determining region 3' (CDR3) sequences were amplified from these samples, from bone marrow samples of 17 other vaccination patients, and from peripheral blood of six healthy controls, cloned and sequenced.We observed a clear bias towards Vβ11 usage of the WT1-specific CTL populations in all four patients. The predominant CDR3β amino acid (AA) sequence of patient 1 was detected in two other patients. CDR3β loops with closely related AA sequences were only found in patient 1. There were no CDR3β AA sequences with side chains of identical chemical properties detected in any patient.We provide the first data addressing TCR Vβ chain usage in WT1-specific T-cell populations after HLA A*0201-restricted single peptide vaccination. We demonstrate both shared Vβ restriction and the sharing of a TCR β transcript with proven clinical impact in one patient.

Agents targeting the mammalian target of rapamycin (mTOR) pathway, e. g. everolimus, can provide clinical benefit in pretreated patients with metastatic renal cell carcinoma (mRCC), but data from randomized trials on the sequential use of temsirolimus are lacking. We retrospectively studied the efficacy and safety of temsirolimus therapy following failure of rTKI therapy.Twenty-nine patients treated with temsirolimus (25 mg/week) following progression on rTKI therapy were studied at four institutions. All patients had failed at least one prior rTKI therapy (sunitinib, n = 6; sorafenib, n = 1; both, n = 22). Over 80% had two or more prior therapies. Data on efficacy (response assessment, progression-free survival [PFS], overall survival [OS]) and safety (NCI-CTC) were analyzed.Adverse events occurred in 90% of patients with the majority being grade 1 (n = 4, 14%) or grade 2 (n = 12, 41%). Most grade 3/4 toxicities (n = 10, 34%) were manageable and included anemia (n = 4, 14%), leukopenia/neutropenia (n = 2, 7%), hyperglycemia (n = 1, 3%), acidosis/alkalosis (n = 2, 7%), and infection (n = 1, 3%). One patient discontinued temsirolimus for grade 3 pneumonitis. Median (range) PFS and OS were 5.1 months (1-10.4) and 18.0 months (12.6-23.3), respectively. Best response included partial response (n = 1) and stable disease (n = 15) for a disease control rate of 55%, and disease progression of 45% (n = 13).Temsirolimus after rTKI failure appears to provide promising safety and efficacy comparable to other treatment options in pretreated patients with mRCC.

Erstmalig seit 3 Jahrzenten stehen neue Medikamente fur das metastasierte Melanom vor der baldigen Zulassung, und die Behandlung wird sich durch Early-Access-Programme bereits ab 2011 deutlich andern. Die Therapie des malignen Melanoms im Stadium der Fernmetastasierung bleibt fur die Mehrzahl der Patienten dennoch palliativ. Moglichst lang andauernde Remissionen mit Verlangerung der Uberlebenszeit werden angestrebt, wobei bei einem kleineren Teil der Patienten auch Heilungen mit den neuen Medikamenten moglich erscheinen. Soweit die Metastasierung ein Organsystem nicht uberschreitet und eine operative Entfernung des Tumors moglich ist, ist die operative Metastasenresektion die Therapie der Wahl. Bei ausgedehnter Metastasierung ist eine Systemtherapie indiziert. Neue Therapiestrategien wie die Blockade von „cytotoxic T lymphocyte-associated antigen 4“ (CTLA-4) durch Antikorper induzierten bei einigen Patienten teils lang anhaltende Tumorremissionen und fuhrten zu einer statistisch evidenten Verlangerung des Gesamtuberlebens. Der therapeutische Einsatz von hoch selektiven Tyrosinkinaseinhibitoren zeigte in ersten Phase-II-Studien objektive Ansprechraten von ca. 80% bei vorigem Nachweis der BRAF-V600E-Mutation (bei 40–50% der Melanompatienten vorhanden) und verdeutlicht uberzeugend die Notwendigkeit einer individualisierten Tumortherapie in der Zukunft.

BACKGROUND: Safety and cellular immunogenicity of rising doses and varying regimens of a poly-epitope vaccine were evaluated in advanced metastatic melanoma. The vaccine comprised plasmid DNA and recombinant modified vaccinia virus Ankara (MVA) both expressing a string (Mel3) of seven HLA.A2/A1 epitopes from five melanoma antigens. METHODS: Forty-one HLA-A2 positive patients with stage III/IV melanoma were enrolled. Patient groups received one or two doses of DNA.Mel3 followed by escalating doses of MVA.Mel3. Immunisations then continued eight weekly in the absence of disease progression. Epitope-specific CD8+ T cell responses were evaluated using ex-vivo tetramer and IFN-gamma ELISPOT assays. Safety and clinical responses were monitored. RESULTS: Prime-boost DNA/MVA induced Melan-A-specific CD8+ T cell responses in 22/31 (71%) patients detected by tetramer assay. ELISPOT detected a response to at least one epitope in 10/31 (32%) patients. T cell responder rates were

Approximately 20% of skin tumors occur in the head and neck region. The therapeutic procedure is laid down in the current interdisciplinary guidelines of the EBM on skin tumors (squamous epithelial carcinoma, melanoma, Merkel cell carcinoma and basal cell carcinoma). For mucous membrane tumors primary R0 surgery is recommended. Approximately 57% of Merkel cell carcinomas occur in the head and neck region. Bilateral selective neck dissection and adjuvant radiation therapy is recommended for every stage. Operative therapy with histological confirmation is the standard procedure for treatment of basal cell carcinoma. The therapeutic procedure for squamous epithelial carcinoma depends on the extent and the histological subtype of the tumor. Complete surgical excision with topographically assigned histopathological control of the incision edges is the first choice treatment as long as this is medically acceptable. Malignant growths in the region of the nasal and paranasal cavities form approximately 4% of head and neck tumors. The general therapeutic principles depend on the possibility of complete resection of the tumor. Postoperative radiation and chemotherapy should be carried out depending on experience with other head and neck tumors in a pT3/4 situation. A dilemma with rare soft tissue sarcomas with many histological subtypes is that at present there is no appropriate classification for this group of tumors. In addition there are no guidelines or evidence with respect to indications for resection of residues after primary chemotherapy of pediatric rhabdomyosarcoma-like tumors. It is therefore recommended that these tumors should be treated in interdisciplinary oncology centers. Small cell carcinomas are highly malignant and rapidly form metastases. The therapy of small cell carcinomas in the head and neck region depend on the localization, tumor spread, lymph node status and the presence of distant metastases.

Dieser Beitrag gibt einen Uberblick zu folgenden „seltenen“ Kopf-Hals-Tumoren: den Speicheldrusenkarzinomen, Plattenepithelkarzinomen, Nasopharynxkarzinomen und Paragangliomen. Aufgrund der Seltenheit der Tumoren liegen in vielen Fallen keine prospektiv-randomisierten Studien vor. Die Empfehlungen zur Therapie beruhen daher uberwiegend auf retrospektiven Studien oder auf den Erfahrungen der Autoren. Der Artikel befasst sich ausschlieslich mit malignen Tumoren. Die Prognose der Speicheldrusenkarzinome richtet sich neben dem histologischen Subtyp auch nach dem Differenzierungsgrad. Die Therapie ist primar chirurgisch und orientiert sich an den bildmorphologischen Kriterien der „sinnvollen“ Resektablilitat. Die Indikation zur adjuvanten Radiotherapie von Speicheldrusenkarzinomen besteht beim Vorliegen von Risikofaktoren, die postoperativ die lokoregionare Rezidivwahrscheinlichkeit erhohen. Plattenepithelkarzinome sind als auserst maligne anzusehen, stellen sich aber vergleichsweise als radio- und chemosensibel dar. Bei Nasopharynxkarzinomen in den fruheren Stadien I–IIA ist die alleinige Radiotherapie, im fortgeschrittenen Stadium III/IV die simultane cisplatinhaltige Radiochemotherapie Standard. Die Wirksamkeit der Chemotherapie ist bei dieser Entitat besonders hoch. Paragangliome sind primar chirurgisch anzugehen. Sie sind grundsatzlich strahlensensibel, aber kaum chemosensibel.

CCR6 is expressed in various tumors and has been implicated in the process of tumor progression and metastasis. Its chemokine ligand, CCL20, is present in different tissues including lymph nodes, but also the normal prostate. This study was performed to investigate a potential relationship between CCR6 and CCL20 expression and features of human prostate cancer (PCA) at time of primary treatment. Immunohistochemistry was used to detect CCR6 and CCL20 expression in archival tissue blocks of 80 PCA cases of various tumor grades and stages. Evaluation was semiquantitatively by visual scoring and quantitatively by digital image analysis (DIA). CCR6 and CCL20 expression was compared with Gleason score, stage, perineural invasion, nodal metastasis, age, and preoperative serum prostate-specific antigen (PSA) level by univariate and multivariate analyses. Staining intensity of CCR6 in tumor cells varied considerably, with it being: weak in 21 tumors (26.2%), intermediate in 44 (55.0%), and strong in 15 (18.8%), with 3.6-log differences in DIA measurements. CCL20 expression was absent in eight tumors (10.0%), weak in 41 (51.2%), intermediate in 23 (28.8%), and strong in eight (10.0%). CCR6 and CCL20 expression did not correlate. CCR6 expression was associated with T-category (P

Recombinant antibody fusion constructs with heterologous functional domains are a promising approach to new therapeutic targeting strategies. However, expression of such constructs is mostly limited to cost and labor-intensive mammalian expression systems. Here we report on the employment of Pichia pastoris for the expression of heterologous antibody fusion constructs with green fluorescent protein, A33scFv::GFP, or with cytosine deaminase, A33scFv::CDy, their production in a biofermenter and a modified purification strategy. Combined, these approaches improved production yields by about thirty times over established standard protocols, with extracellular secretion of the fusion construct reaching 12.0 mg/l. Bifunctional activity of the fusion proteins was demonstrated by flow cytometry and an in-vitro cytotoxicity assay. With equal amounts of purified protein, the modified purification method lead to higher functional results. Our results demonstrate the suitability of methylotrophic Pichia expression systems and laboratory-scale bioreactors for the production of high quantities of bifunctionally active heterologous single-chain fusion proteins.

In recent years, reduced-intensity conditioning (RIC) regimens before allogeneic stem cell transplantation (SCT) are increasingly used in patients not eligible for conventional conditioning. We did a retrospective, multicenter analysis to assess the feasibility of conditioning with fludarabine and treosulfan before allogeneic SCT in multiple myeloma patients. Thirty-four patients with a median age of 51.5 years were included in the analysis. All patients underwent myeloablation after conditioning followed by stable engraftment, and 29 of 31 evaluable patients (94%) showed early complete hematopoietic chimerism. Non-hematological toxicities were limited and encompassed mainly fever in neutropenia and infections. Grade II-IV acute and chronic graft-versus-host disease was observed in 33 and 39%, respectively. With a median follow-up of 708 days (range 60-1729 days), the median progression-free survival was 180 days. The treatment-related mortality was 10% on day 100 and 25% after 1 year. The median overall survival has not yet been reached. Our data indicate that conditioning with fludarabine and treosulfan before allogeneic SCT is feasible in intensively pretreated multiple myeloma patients and leads to stable engraftment and complete hematopoietic chimerism. Randomized trials are warranted to determine if this approach might be incorporated in an algorithm of multiple myeloma treatment.

Die palliative medikamentose Systemtherapie von Patienten mit Plattenepithelkarzinomen der Kopf-Hals-Region ist ein komplexes Feld, zu dem nur wenige Daten vorliegen und das von zahlreichen Einzelfallentscheidungen gepragt ist. Eine Reihe zytostatischer Substanzen ist fur die Behandlung zugelassen, z. B Platinsalze, 5-Fluorouracil, Methotrexat. Die Taxane gewinnen derzeit zunehmend an Bedeutung. Zusatzlich besteht die Moglichkeit der EGF-Rezeptorblockade mittels Antikorpern und Tyrosinkinaseinhibitoren. Der Stellenwert der Behandlung mit angiogeneseinhibierenden Substanzen ist noch unklar. Da eine Kombinationsbehandlung in der palliativen Therapie keinen eindeutigen Vorteil zeigt, werden die verfugbaren Medikamente in der Regel sequenziell eingesetzt, immer unter Beachtung der bei dieser spezifischen Patientengruppe haufigen Komorbiditaten.

Organerhaltende Therapieansatze bei Karzinomen des Larynx und Hypopharynx als Alternative zur kompletten Kehlkopfentfernung (Laryngektomie) wurden in den letzten 2 Jahrzehnten signifikant weiterentwickelt und in der klinischen Betrachtung etabliert. Insbesondere die transorale Laserchirurgie hat sich einen festen Stellenwert in der Behandlung mikrolaryngoskopisch gut ubersehbarer Karzinome (bis Tumorgrose T3) erarbeitet, wobei je nach Erfahrung des Operateurs in Einzelfallen auch offene Teilresektionen des Kehlkopfs zur Anwendung kommen. Ubereinkunft herrscht bezuglich der nach wie vor gultigen Notwendigkeit der Laryngektomie, die bei fortgeschrittenen Tumoren des Larynx und Hypopharynx oft die rehabilitativ gunstigste und onkologisch sauberste Therapie der Wahl darstellt. Hat das Karzinom eine Ausbreitung, die durch Teilresektionen nicht mehr sinnvoll behandelbar ist, konnen multimodale Therapieansatze zum Einsatz kommen, die in den letzten Jahren im Rahmen klinischer Studien diversifiziert wurden. Fruh konnte gezeigt werden, dass solche Organerhaltungsprogramme in onkologischer Hinsicht gleichwertig mit der Laryngektomie zu setzen sind und langfristig funktionell zufrieden stellende Organerhaltungsraten bis zu 60% erreichen konnen. Der aktuelle Fokus richtet sich derzeit auf den Stellenwert der Induktionschemotherapie, dem zusatzlichen Einsatz von Biologicals (insb. EGFR-Blockade) und die Reduktion der Spattoxizitat.

Das Aderhautmelanom unterscheidet sich in der Tumorbiologie und im Metastasierungsverhalten vom kutanen malignen Melanom. Das metastasierte Aderhautmelanom ist eine seltene Erkrankung mit schlechter Prognose. Zu den therapeutischen Moglichkeiten zahlen operative Verfahren, die nur fur sehr wenige Patienten sinnvoll sind, die systemische Chemotherapie und lokale Behandlungsverfahren wie die Chemoembolisation und die lokoregionare Chemotherapie uber die A. hepatica. Die wenigen vorliegenden klinischen Studien haben nur eine geringe Zahl von Patienten eingeschlossen. Sie belegen eine Wirksamkeit der Chemoembolisation und der lokoregionaren Chemotherapie bei Patienten mit alleiniger Metastasierung in die Leber. Bei Patienten mit Metastasierung in mehrere Organe werden zurzeit Zytostatikakombinationen aus Gemcitabin und Treosulfan in klinischen Studien untersucht. Fortemustin, eine moderne Nitroseharnstoffverbindung, stellt ebenfalls eine therapeutische Option dar.

Die Behandlungsmodalitat des primaren Aderhautmelanoms hat bisher keinen direkten Einfluss auf die Uberlebensprognose des Patienten. Durch die Identifizierung signifikanter Prognoseparameter bezuglich des Risikos, Metastasen eines Aderhautmelanoms zu entwickeln, werden neue Behandlungsstrategien erarbeitet mit dem Ziel, die Entstehung von Metastasen zu verhindern. Dabei spielt die Immuntherapie durch die Entwicklung geeigneter Impfstoffe gegen melanomspezifische Tumorantigene eine zunehmende Rolle. Beim bereits metastasierten Aderhautmelanom betrug bis dato die mittlere Uberlebenszeit ca. 5 Monate. In der letzten Zeit konnte durch die intrahepatische Infusion von Fotemustin die mittlere Uberlebenszeit auf 14 Monate bei selektierten Patienten verlangert werden. Neue Therapieprotokolle werden aufgrund von Chemosensitivitatsuntersuchungen aktuell erarbeitet. Des Weiteren werden immuntherapeutische Modalitaten auch in der Behandlung von Metastasen von Aderhautmelanomen untersucht.

Integrins are cell surface molecules that mediate cell adhesion, but are also important regulators of tumor cell interactions with their microenvironment, tumor cell survival and growth. In addition, the αvβ3-integrins appear to be critical for microvessel formation in tumor-induced neoangiogenesis. The present study is the first to investigate the effects of therapeutic αvβ3-integrin inhibition in a chemically induced tumor model that largely resembles human colon carcinomas. Tumor induction was performed in 47 male Sprague-Dawley rats using 1,2 dimethylhydrazin (21 mg/kg) twice a week. After 20 weeks of tumor induction, 100% of the animals developed adenocarcinomas with a median of 13.5 macroscopic tumor nodules (range 12–17), but no distant metastases. During further tumor induction for an additional 10 weeks, rats were treated three times/week with (a) 15 mg/kg RGDfV-peptide that can block vitronectin and fibronectin receptors; (b) an equimolar amount of an ineffective cyclic control peptide; or (c) with equimolar amounts of a linear RGDS-peptide. At the end of this treatment period, rats were sacrificed, and tumor load was quantified macroscopically and confirmed by histological examination. For investigation of the involvement of tumor-induced neoangiogenesis microvessel, density was determined using CD31-immunostaining. After 30 weeks, control animals (group B) had 5–18 tumors (median 14.5). If rats were treated with RGDfV-peptide (group A), the number of tumor nodules was significantly reduced (P < 0.005) to a median of seven macroscopic tumors (range 2–10 tumors), which also represented a significant reduction (P < 0.005) compared with prior to treatment. Application of noncylic RGDS-peptides (group C) did not affect the number of tumor nodules (median 18; range 10–30 tumors). The diameters of tumor nodules were comparable (3.2–6.1 mm) in animals of all groups. In addition, microvessel density was significantly (P < 0.05) reduced in tumors in group A compared to control rats. The major side effect in the treatment group was increased susceptibility to respiratory infections. Our results demonstrate that αvβ3-integrin-receptor inhibition appears to be a therapeutic strategy for colorectal cancer. In our therapeutic model, late onset of treatment with integrin-blocking peptides resulted in an inhibition of tumor growth and a reduced tumor load which appeared to be mediated, at least in part, by inhibition of neoangiogenesis.

The clinical value of the reverse transcription polymerase chain reaction (RT-PCR) assay for tyrosinase in peripheral blood of melanoma patients is still under debate. A total of 212 blood samples from 212 melanoma patients in all clinical stages (AJCC) were examined. Erythrocytes were lysed prior to RNA extraction by phenol precipitation from 2.7 ml of blood. cDNA for tyrosinase PCR was synthesized using random hexamers. Positive tyrosinase RT-PCR results were obtained in 11% of 106 stage I patients, 18% of 56 stage II patients, 31% of 26 stage III patients and 67% of 24 stage IV patients. After a median follow-up of 36 months (range 26–41), stage III patients with positive RT-PCR for tyrosinase had a shortened disease-free interval as compared to negative patients (P< 0.01). In stage IV patients, median overall survival was 8 months in case of a positive RT-PCR in contrast to 12 months in case of a negative test. While univariate analysis showed sex and primary tumour location associated with positive RT-PCR, multiple regression analysis revealed clinical stage and detection of tyrosinase transcripts in peripheral blood as best prognostic factors. Hazard ratios for disease-free survival were 19.7 (confidence interval (CI) 8.53–45.5, P = 0.0001) for metastatic vs primary disease and 2.96 (Cl 1.49–5.89, P = 0.002) for positive vs negative tyrosinase RT-PCR. The corresponding hazard ratios for overall survival were 97.0 (Cl 12.7–741, P = 0.0001) and 4.33 (Cl 1.69–11.1, P = 0.002). Our results emphasize the importance of tyrosinase RT-PCR testing in peripheral blood. © 2000 Cancer Research Campaign

Clonal expansions of T cells carrying identical T-cell-receptor (TCR) genes are the hallmark of T-cell malignancies, but they can also result from a strong immune reaction to a dominant epitope. The basis for the molecular detection of clonal T cells is amplification of the V-(D)-N-J region of the TCR gene. We evaluated PCR amplification of the rearranged gamma TCR from genomic DNA extracted from peripheral blood and subsequent polyacrylamide gel electrophoresis (PAGE) in an automated DNA sequencer. We determined the sensitivity for the detection of clonal T cells and propose a standardized evaluation procedure for the electrophoretic profiles generated by the DNA sequencer. The sensitivity of our method was 0.6-1.25% of clonal T cells within a polyclonal background. Sixteen patients with T-cell malignancies, ten with acute inflammatory rheumatic diseases, and twelve healthy controls were examined. Among the systemic T-cell malignancies, all but one patient with T-PLL (8/ 9) revealed a clonal PCR signal. No clonal signal was detectable in any patient in clinical complete remission (5/5) or in either of the two patients with lymphomas limited to cutaneous sites. However, clonal T cells were detected in one patient with polymyalgia rheumatica and in one with reactive arthritis. A polyclonal signal was found in the remaining eight patients with acute inflammatory rheumatic diseases and in 12 healthy controls. Taking our results together, the PCR/PAGE assay is able to reliably distinguish clonal from polyclonal T-cell populations. However, although the sensitivity is limited to approximately 1%, clonal T cells can be found in the peripheral blood of some patients with autoimmune diseases and not only in T-cell malignancies.

Background A limitation of positive selection strategies to enrich for circulating tumor cells (CTCs) is that there might be CTCs with insufficient expression of the surface target marker which may be missed by the procedure. We optimized a method for enrichment, subsequent detection and characterization of CTCs based on depletion of the leukocyte fraction. Methods The 2-step protocol was developed for processing 20 mL blood and based on red blood cell lysis followed by leukocyte depletion. The remaining material was stained with the epithelial markers EpCAM and cytokeratin (CK) 7/8 or for the melanoma marker HMW-MAA/MCSP. CTCs were detected by flow cytometry. CTCs enriched from blood of patients with carcinoma were defined as EpCAM+CK+CD45-. CTCs enriched from blood of patients with melanoma were defined as MCSP+CD45-. One-hundred-sixteen consecutive blood samples from 70 patients with metastatic carcinomas (n = 48) or metastatic melanoma (n = 22) were analyzed. Results CTCs were detected in 47 of 84 blood samples (56%) drawn from carcinoma patients, and in 17 of 32 samples (53%) from melanoma patients. CD45-EpCAM-CK+ was detected in pleural effusion specimens, as well as in peripheral blood samples of patients with NSCLC. EpCAM-CK+ cells have been successfully cultured and passaged longer than six months suggesting their neoplastic origin. This was confirmed by CGH. By defining CTCs in carcinoma patients as CD45-CK+ and/or EpCAM+, the detection rate increased to 73% (61/84). Conclusion Enriching CTCs using CD45 depletion allowed for detection of epithelial cancer cells not displaying the classical phenotype. This potentially leads to a more accurate estimation of the number of CTCs. If detection of CTCs without a classical epithelial phenotype has clinical relevance need to be determined.

Der Fortschritt in der systemischen Behandlung von Patienten mit metastasiertem Melanom war uber die letzten 25 Jahre sehr begrenzt. Erst in den vergangenen 10–15 Jahren wurden eine Reihe randomisierter Studien durchgefuhrt, die eine Bewertung der zur Verfugung stehenden Medikamente im Sinne evidenzbasierter Medizin ermoglichen. In Deutschland sind die Zytostatika Dacarbazin, Vindesin und Cisplatin zugelassen, in anderen Industrienationen auch Fotemustin und das Zytokin Interleukin-2. Prinzipielles Fazit der randomisierten Studien ist, dass Kombinationsprotokolle mit mehreren Zytostatika bzw. Zytostatika-Zytokinkombinationen entgegen den ursprunglichen Hoffnungen keinen Uberlebensvorteil bringen, sodass die Empfehlungen wieder dahin gehen, Einzelsubstanzen einzusetzen und Patienten bevorzugt im Rahmen klinischer Studien zu behandeln. Ausnahme ist die Behandlung von Patienten mit erheblicher tumorbedingter Symptomatik, die den Einsatz einer Polychemotherapie mit palliativer Zielsetzung unter Inkaufnahme erhohter Toxizitat rechtfertigt. Im Rahmen verschiedener klinischer Zulassungsstudien wurden die Kombination aus Histamin und Interleukin-2 sowie von Dacarbazin mit Bcl-2-Antisense-Oligonukleotid gepruft. Ergebnisse und evtl. zugar eine Zulassung in den nachsten 12 Monaten konnten die Folge sein.

Complete remission in a patient with recurrent acute myeloid leukemia induced by vaccination with WT1 peptide in the absence of hematological or renal toxicity

Introduction Spontaneous T cell responses against specific tumor-associated antigens (TAA) are frequently detected in peripheral blood of tumor patients of various histiotypes. However, little is known about whether these circulating, spontaneously occurring, TAA-reactive T cells influence the clinical course of disease.