[Durable remission under dual HER2 blockade with Trastuzumab and Pertuzumab in a patient with metastatic gallbladder cancer]. / Dauerhafte Remission unter dualer HER2-Blockade mit Trastuzumab und Pertuzumab bei metastasiertem Gallenblasenkarzinom.

Gallbladder cancer represents a rare but dismal disease. The only curative option is complete surgical resection, though patients often develop recurrent disease. In patients with advanced biliary tract cancer, the combination of cisplatin and gemcitabine showed a benefit in overall survival compared to gemcitabine alone. However, there is no standardized second-line regimen after treatment failure. We report on a young patient with early recurrence of a gallbladder cancer with cutaneous and peritoneal metastases. Upon identification of an ERBB2 gene amplification within the NCT MASTER (Molecularly Aided Stratification for Tumor Eradication Research) exome sequencing program with resulting overexpression of HER2 in the tumors cells, the patient received a targeted therapy with the HER2 antibodies pertuzumab and trastuzumab in combination with nab-paclitaxel, which led to a durable remission for more than one year. This case report underlines the potential of molecularly aided personalized targeted therapy for patients with biliary tract cancer and the need for respective clinical trials.

Targeted lentiviral vectors pseudotyped with the Tupaia paramyxovirus glycoproteins.

Lentiviral vectors are vectors of choice for many gene therapy applications. Recently, efficient targeting of lentiviral vectors pseudotyped with the Measles virus (MV) glycoproteins has been reported. However, MV antibodies in patients might limit the clinical use of these vectors. We demonstrate here that lentiviral vectors can also be pseudotyped with the glycoproteins of Tupaia paramyxovirus (TPMV), the hemagglutinin (H) and fusion (F) protein. As this animal paramyxovirus has no known close relatives in humans, we do not expect TPMV antibodies in patients. Because TPMV normally does not infect human cells, 'detargeting' from natural receptors is unnecessary. Similar to the MV system, TPMV glycoproteins can mediate targeted cell entry by displaying different single-chain antibodies (scAb) directed against surface molecules on target cells on the viral hemagglutinin. We generated a panel of H and F proteins with truncated cytoplasmic tails and determined the variants that efficiently pseudotyped lentiviral vectors. The B-cell marker CD20 was used as a model antigen, and CD20-targeted TPMV vectors selectively transduced CD20-positive cells, including quiescent primary human B-cells. Lentiviral vectors pseudotyped with targeted TPMV envelope proteins might be a valuable vector choice when systemic application of targeted lentiviral vectors in humans is required.

[Neoadjuvant and adjuvant treatment of pancreatic cancer]. / Neoadjuvante und adjuvante Therapie beim Pankreaskarzinom.

BACKGROUND: Resection is the only curative treatment option for pancreatic cancer patients. Adjuvant chemotherapy can improve disease-free survival and overall survival after resection. Furthermore, neoadjuvant treatment protocols are currently being investigated in a large number of studies. OBJECTIVE: Summary of the current evidence for adjuvant and neoadjuvant treatment. MATERIAL AND METHODS: Review of the current scientific literature and guidelines. RESULTS AND CONCLUSION: After resection for pancreatic cancer patients should receive intensive chemotherapy with mFOLFIRINOX, gemcitabine plus capecitabine or gemcitabine/5-fluorouracil (5-FU) monotherapy. Neoadjuvant treatment concepts are promising but have to be further evaluated in prospective studies.

Fatal varicella in an immunocompromised adult associated with a European genotype E2 variant of varicella zoster virus.

Varicella zoster virus (VZV) seronegative patients under immunosuppressive therapy are at risk for severe life-threatening varicella. A 25-year-old male patient presented with rash and hepatitis. He had been known to suffer from Crohn's disease and received immunosuppressive treatment with azathioprine. The patient showed dyspnoea, and presented with a generalized rash with vesicular lesions typical for herpesvirus infections. He was started immediately on acyclovir therapy. Varicella infection was determined in this VZV seronegative patient in rash vesicles, blood and tracheal secretions and a high VZV viral load was detected in these specimens. The causative agent was genotyped by sequencing of several genes as a variant of the European genotype E2 containing several unique single nucleotide polymorphisms. Despite all measures, there was progressive septic shock, and the patient died due to multi-organ failure. Immunocompromised adults without varicella history are at high risk to develop life-threatening complications of varicella. Antiviral therapy with acyclovir can only be successful when administered as early as possible on suspicion of varicella infection in this group of patients. The most effective method to prevent severe varicella in immunocompromised patients is active immunization prior to immunosuppressive therapy.

Surgery in oesophago-gastric cancer with metastatic disease: Treatment, prognosis and preoperative patient selection.

BACKGROUND: The role of surgical resection in metastatic oesophago-gastric adenocarcinomas (EGA) is not defined and regularly discussed in interdisciplinary tumour boards. Primary objective of this retrospective study was the outcome of patients after surgery. We additionally evaluated our preoperative prognostic score (PPS) based on tumour grading, clinical response to chemotherapy and presumed R-status. METHODS: 123 of 811 EGA patients were evaluated as cM1, either confirmed intraoperatively or by imaging. Response evaluation after chemotherapy was performed by endoscopy, CT-scan and histopathologically. The prospectively documented patient and outcome data were analysed retrospectively. RESULTS: 70 patients with adenocarcinoma of the oesophago-gastric junction and 53 patients with gastric cancer were included. The majority had one M1 site (n = 102). 72 received preoperative chemotherapy (CTx) and 51 underwent primary resection. 11 were explored without resection. 49/112 (40%) had multivisceral resections and 63/112 (56%) were completely resected (R0). 26/72 (36%) were clinical responders and 30 patients had a favourable PPS. Median survival was 20.0 months. Survival was significantly prolonged by resection, especially complete resection, and by preoperative CTx (all p = 0.001). Multivisceral resection, type or number of metastases, or primary tumour localization had no impact on survival. In patients undergoing preoperative CTx, clinical response and the PPS influenced survival significantly. In R0 resected patients, preoperative CTx, clinical response and the PPS remained prognostic. CONCLUSION: Primary resection without preoperative CTx is not appropriate for metastatic EGA. Subgroups of patients with a favourable PPS with response to CTx may be good candidates for surgical resection in metastatic oesophago-gastric cancer.

Structural organization of a conserved gene cluster of Tupaia herpesvirus encoding the DNA polymerase, glycoprotein B, a probable processing and transport protein, and the major DNA binding protein.

The Tupaia herpesviruses (THVs) have been isolated from malignant lymphoma tissue cultures and from degenerating lung and spleen cell cultures of tree shrews (Tupaia spp.). Recently we succeeded in the localization of the gene locus of the THV DNA polymerase (DPOL) gene within the viral genome. Based on these results the highly conserved gene cluster of herpesviruses encoding the DPOL, the glycoprotein B (gB), a probable processing and transport protein (PRTP), and the major DNA binding protein (DNBI) was characterized in the genome of THV strain 2 (THV-2) in its entirety. The complete nucleotide sequence of the gene cluster was determined and it was discovered that the THV-2 gene products are most closely related to the corresponding proteins of mammalian cytomegaloviruses. The transcriptional activity of the four genes was confirmed by amplification of a part of the corresponding mRNAs obtained from infected cell RNA by RT-PCR. The homology values and the overall structure of the gene cluster, that shows specific colinearity with the corresponding clusters of the mammalian cytomegaloviruses, is further evidence that THV-2 is a member of the subfamily Betaherpesvirinae.

Chemovirotherapy for head and neck squamous cell carcinoma with EGFR-targeted and CD/UPRT-armed oncolytic measles virus.

First-line treatment of recurrent and/or refractory head and neck squamous cell carcinoma (HNSCC) is based on platinum, 5-fluorouracil (5-FU) and the monoclonal antiEGFR antibody cetuximab. However, in most cases this chemoimmunotherapy does not cure the disease, and more than 50% of HNSCC patients are dying because of local recurrence of the tumors. In the majority of cases, HNSCC overexpress the epidermal growth factor receptor (EGFR), and its presence is associated with a poor outcome. In this study, we engineered an EGFR-targeted oncolytic measles virus (MV), armed with the bifunctional enzyme cytosine deaminase/uracil phosphoribosyltransferase (CD/UPRT). CD/UPRT converts 5-fluorocytosine (5-FC) into the chemotherapeutic 5-FU, a mainstay of HNSCC chemotherapy. This virus efficiently replicates in and lyses primary HNSCC cells in vitro. Arming with CD/UPRT mediates efficient prodrug activation with high bystander killing of non-infected tumor cells. In mice bearing primary HNSCC xenografts, intratumoral administration of MV-antiEGFR resulted in statistically significant tumor growth delay and prolongation of survival. Importantly, combination with 5-FC is superior to virus-only treatment leading to significant tumor growth inhibition. Thus, chemovirotherapy with EGFR-targeted and CD/UPRT-armed MV is highly efficacious in preclinical settings with direct translational implications for a planned Phase I clinical trial of MV for locoregional treatment of HNSCC.

TT virus as a human pathogen: significance and problems.

In 1997 TTV was detected using representational difference analysis (RDA) in serum of a patient with posttransfusion hepatitis unrelated to known hepatitis viruses. The genome of TTV is a circular single-stranded DNA molecule of 3852 nt with negative polarity. TTV possibly can be grouped either into the existing family Circoviridae or into a recently established virus family "Circinoviridae". Analysis of the complete DNA nucleotide sequence of TTV identified three partially overlapping open reading frames (ORFs). Neither DNA nucleotide nor corresponding amino acid sequences of TTV do show significant homologies to known sequences. TTV DNA nucleotide sequences amplified by PCR from sera of different patients show considerable sequence variations. Although the natural route of transmission of TTV is still unknown, there is clear evidence for a transmission of TTV through blood and blood products. TTV DNA can be detected in the feces of infected individuals suggesting that it may be possible to attract TTV infection from environmental sources. Since the discovery of TTV, numerous studies have investigated the prevalence of TTV infections in different human population groups all over the world. All these studies are based on PCR detection systems, but the technical aspects of the PCR systems vary significantly between the different investigators. The results of the epidemiological studies do not show a clear picture. The discovery of TTV as a viral agent and particularly the identification of a high percentage of infected carriers in the healthy human population raises the following questions: Firstly, what is the origin and molecular relatedness of TT virus. Secondly, what is the significance of TTV as a human pathogen. And thirdly, what are the exact molecular mechanisms of viral replication. To answer these questions it will be necessary to determine the primary structure and the coding capacity of several TTV patient isolates.

Identification and characterization of the Tupaia herpesvirus DNA polymerase gene.

Tupaia herpesviruses (THVs) have been isolated from malignant lymphomas and from degenerating lung or spleen cell cultures of tree shrews (Tupoia spp.), but because of a lack of genetic information the final classification of THVs is still open. In the present work the viral DNA polymerase (DPOL) gene was mapped within the genome of the different THV strains using PCR and degenerate oligonucleotide primers. Nucleotide sequences of the DPOL genes of THV strains 1 to 5 were determined and used for comparative analyses. The transcriptional activity of the THV-2 DPOL gene was confirmed by RT-PCR. It was found that the different THV strains are very closely related to each other. When compared to other herpesviruses the highest amino acid sequence identities detected were with DPOLs of the murine and human cytomegaloviruses. These results justify the conclusion that THVs are members of the subfamily Betaherpes-virinae.