Improving therapeutic efficacy of IL-12 intratumoral gene electrotransfer through novel plasmid design and modified parameters.

The use of immunomodulatory cytokines has been shown effective in regressing a wide range of tumors. However, systemic delivery of recombinant cytokines results in serious, potentially life-threatening, adverse effects. By contrast, nucleic acid transfer via electroporation (EP) is a safe and effective method of delivering plasmid-encoded cytokines to tumors. Intratumoral delivery of IL-12 plasmid DNA by electroporation (IT-pIL12-EP) produced objective response rates in Phase 2 clinical trials in metastatic melanoma. However, only 17.9% of patients receiving IT-pIL12-EP show a complete therapeutic response. Here, we sought to improve the antitumor efficacy of our clinical IT-pIL12-EP plasmid electroporation platform. We evaluated multiple plasmid designs for IL-12 expression. IL-12 expression from a plasmid incorporating a picornavirus-derived co-translational P2A site was the most effective in expressing IL-12p70. In addition, modifying the electroporation parameters improved transfection efficiency and expression of plasmid-derived IL-12p70, as well as its downstream effector IFN-γ in vivo. Finally, using a murine melanoma model that is representative of the intended target patient population, we show that combining modified electroporation conditions with the pIL12-P2A plasmid expression enhances the systemic antitumor response. These improvements to the IT-pIL12-EP platform may improve patient clinical response rates and survival when translated to clinical trials.

Contribution of increased ISG15, ISGylation and deregulated type I IFN signaling in Usp18 mutant mice during the course of bacterial infections.

Host genetics has a key role in susceptibility to Salmonella Typhimurium infection. We previously used N-ethyl-N-nitrosourea (ENU) mutagenesis to identify a loss-of-function mutation within the gene ubiquitin-specific peptidase 18 (Usp18(Ity9)), which confers increased susceptibility to Salmonella Typhimurium. USP18 functions to regulate type I interferon (IFN) signaling and as a protease to remove ISG15 from substrate proteins. Usp18(Ity9) mice are susceptible to infection with Salmonella Typhimurium and have increased expression and function of ISG15, but Usp18(Ity9) mice lacking Isg15 do not show improved survival with Salmonella challenge. Type I IFN signaling is increased in Usp18(Ity9) mice and inhibition of type I IFN signaling is associated with improved survival in mutant mice. Hyperactivation of type I IFN signaling leads to increased IL-10, deregulated expression of autophagy markers and elevated interleukin (IL)-1ß and IL-17. Furthermore, Usp18(Ity9) mice are more susceptible to infection with Mycobacterium tuberculosis, have increased bacterial load in the lung and spleen, elevated inflammatory cytokines and more severe lung pathology. These findings demonstrate that regulation of type I IFN signaling is the predominant mechanism affecting the susceptibility of Usp18(Ity9) mice to Salmonella infection and that hyperactivation of signaling leads to increased IL-10, deregulation of autophagic markers and increased proinflammatory cytokine production.

[Intratympanic steroid administration: use in the treatment of profound idiopathic sudden sensorineural hearing loss]. / Intratympanale Dexamethasongabe: Nutzen in der Behandlung des hochgradigen Hörsturzes.

BACKGROUND: Therapy for idiopathic sudden sensorineural hearing loss is still controversial. Although there are no evidenced-based studies, therapy with systemic steroids is widely accepted as the gold standard. Intratympanic administration of steroids appears to be an alternative or additional method of management without the disadvantage of systemic side effects and, therefore, makes therapy accessible for patients with contraindication for systemic steroids. MATERIAL AND METHODS: This retrospective analysis compares the audiometric results of 25 patients who were treated with standard therapy (prednisolone, hydroxyethyl starch, pentoxyfylline) with 23 patients who additionally received intratympanic steroids (IT group). A total of 4 injections were administered within 10 days. The solution used consisted of 0.3 ml dexamethasone (8 mg/ml) and 0.2 ml hyaluronic acid 0.2%. The pure-tone average (PTA) was evaluated prior to and 3 months after treatment. RESULTS: The PTA 3 months after treatment showed an improvement of 48 dB in the IT group and 38 dB in the standard treatment group. The IT group achieved better recovery with an average PTA improvement of 68% compared to the standard treatment group with an average improvement of 59%. Neither result reached significance. CONCLUSION: Combination therapy with intratympanic steroids showed a tendency for better hearing results without serious side effects. However, because current evidence is not adequate, randomized placebo-controlled multicenter studies are needed.

Paclitaxel and carboplatin vs gemcitabine and vinorelbine in patients with adeno- or undifferentiated carcinoma of unknown primary: a randomised prospective phase II trial.

Platinum/taxane combinations are widely used in patients with carcinoma of unknown primary (CUP), yielding response rates of 30% and median overall survival of 9-11 months in selected patients. Yet these combinations have not been subject to a randomised trial to overcome selection bias, a major problem in CUP. We randomised 92 patients to either paclitaxel/carboplatin (arm A) or the non-platinum non-taxane regimen gemcitabine/vinorelbine (arm B). The primary endpoint was rate of practicability as defined: application of >or=2 cycles of therapy (1) with a maximal delay of 1 week (2) and survival of >or=8 months (3). Practicability was shown in 52.4% (95% CI 36-68%) in arm A and in 42.2% (95% CI 28-58%) in arm B, respectively. The median overall survival, 1-year survival -rate and response rate of patients treated in arm A was 11.0 months, 38, and 23.8%, arm B 7.0 months, 29, and 20%. In conclusion, the paclitaxel/carboplatin regimen showed clinically meaningful activity in this randomised trial (Clinical trial registration number 219, 'Deutsches KrebsStudienRegister', German Cancer Society.)

Dual-phase multidetector thin-section CT in detecting duodenal gastrinoma.

Preoperative localization of gastrinomas, especially of extrapancreatic origin, remains a challenge to the radiologist. Most patients with extrapancreatic gastrinomas undergo surgery without preoperative identification of the primary tumor. The appropriate imaging modality to localize gastrinomas is under continuing debate. We report a case of a duodenal gastrinoma with regional lymph node metastases that presented with Zollinger-Ellison syndrome. The small primary tumor was detected noninvasively by dual-phase multidetector thin-section computed tomography with adequate bowel distention and confirmed by endoscopy and histopathologic examination. The case illustrates that appropriate computed tomographic technique and scanning protocol are crucial for success in localizing extrapancreatic gastrinoma.

[Ultrasound endometrium follow-up during tamoxifen treatment: Really not reliable or useful after all?]. / Sonographische Endometriumkontrolle unter Tamoxifen: Wirklich nicht empfehlenswert oder doch sinnvoll?

AIM: To investigate whether an examination of the endometrium of women treated with tamoxifen (TAM) is useful or not. METHOD: 40 breast cancer patients who displayed a thickened endometrium of > 8 mm and/or vaginal bleeding were included in the study. They received daily TAM adjuvantly. Histologic clarification by hysteroscopy and D&C was recommended for patients with an endometrium of > 8 mm or vaginal bleeding. RESULTS: In our collective, the mean endometrial thickness was 13.7 +/- 5.6 mm (SD). 32 patients underwent a histological examination. Most had a benign lesion; in 2 cases we merely found a cystic atrophy (11 mm, 18 mm), 2 displayed atypical tissue (13 mm, 25 mm) and 2 an endometrial cancer (19 mm, 33 mm). All patients with atypical tissue or cancer had an endometrial thickness markedly above the norm, but 3 of them were not bleeding. No linear correlation between thickness of the endometrium and duration of TAM intake was found. CONCLUSION: To detect early premalignant or malignant changes of the endometrium, we recommend histological examination by hysteroscopy and dilatation and curettage when the endometrium is > 8 mm thick, even in the absence of symptoms. Therefore, these patients should have regular examinations by transvaginal ultrasound once or twice a year. Moreover, continuing regular screening of the endometrium for years after termination of tamoxifen-therapy is also to be recommended.

[Radiofrequency ablation of liver metastases]. / Radiofrequenzablation von Lebermetastasen.

The liver is the second only to lymph nodes as the most common site of metastatic disease irrespective of the primary tumor. Up to 50% of all patients with malignant diseases will develop liver metastases with a significant morbidity and mortality. Although the surgical resection leads to an improvement of the survival time, only approximately 20% of the patients are eligible for surgical intervention. Radiofrequency (RF) ablation represents one of the most important alternatives as well as complementary methods for the therapy of liver metastases. RF ablation can lead in a selected patient group to a palliation or to an increased life expectancy. RF ablation appears either safer (vs. cryotherapy) or easier (vs. laser) or more effective (percutaneous ethanol instillation [PEI], transarterial chemoembolisation [TACE]) in comparison with other minimal invasive procedures. RF ablation can be performed percutaneously, laparoscopically or intraoperatively and may be combined with chemotherapy as well as with surgical resection. Permanent technical improvements of RF systems, a better understanding of the underlying electrophysiological principles and an interdisciplinary approach will lead to a prognosis improvement in patients with liver metastases.