• Immuno-gene therapy with interferon-beta before surgical debulking delays recurrence and improves survival in a murine model of malignant mesothelioma. (1/2004)

• Regression of AK7 malignant mesothelioma established in immunocompetent mice following intratumoral gene transfer of interferon gamma. (6/2003)

• Analysis of the immunologic response generated by Ad.IFN-beta during successful intraperitoneal tumor gene therapy. (8/2002)

• Intrapleural infusion of activated macrophages and gamma-interferon in malignant pleural mesothelioma: a phase II study (6/2002)

• Combined regimen of cisplatin, doxorubicin, and alpha-2b interferon in the treatment of advanced malignant pleural mesothelioma: a Phase II multicenter trial of the Italian Group on Rare Tumors (GITR) and the Italian Lung Cancer Task Force (FONICAP). (8/2001)

• Control of cell cycle progression in human mesothelioma cells treated with gamma interferon. (3/2001)

Immuno-gene therapy with interferon-beta before surgical debulking delays recurrence and improves survival in a murine model of malignant mesothelioma. (1/2004)

Kruklitis RJ, Singhal S, Delong P, Kapoor V, Sterman DH, Kaiser LR, Albelda SM.

Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, USA.

OBJECTIVES: Immuno-gene therapy of mesothelioma with an adenovirus encoding interferon-beta mediated strong antitumor responses in murine models with low but not high tumor burden. Our goals were to determine the mechanisms responsible for this loss of efficacy and to test the hypothesis that the combination of preoperative adenovirus encoding interferon-beta and surgical resection would be effective in treating bulky tumors.

METHODS: Flank tumors of a mouse mesothelioma cell line were treated with adenovirus encoding interferon-beta or adenoviral vector encoding the bacterial protein beta-galactosidase. Cytotoxic T lymphocytes and tumor infiltration by T lymphocytes were measured. Tumors were surgically excised 72 hours later and tumor cells were injected in the contralateral flank to create a model of a metastatic focus. Tumor-free survival and distant metastatic disease were assessed.

RESULTS: Immuno-gene therapy effectively treated small tumors (<200 mm(3)) but did not reduce the size of large (>800 mm(3)) flank tumors. Although treatment with adenovirus encoding interferon-beta resulted in the generation of tumor-neutralizing splenocytes in large tumors, the number of T cells visualized within the tumors was minimal. Tumors treated with adenovirus encoding interferon-beta (versus adenoviral vector encoding the bacterial protein beta-galactosidase or phosphate-buffered saline solution) prior to debulking increased long-term tumor-free survival and resulted in two- to sixfold smaller foci of implanted tumor cells at 2 weeks postoperatively.

CONCLUSIONS: The use of adenovirus encoding interferon-beta or surgical debulking alone is ineffective in treating large tumors, but combining preoperative adenovirus encoding interferon-beta and surgical debulking significantly reduces tumor recurrence and improves long-term tumor-free survival. We postulate that adenovirus encoding interferon-beta amplifies the cytotoxic T-lymphocyte antitumor response, allowing elimination of residual tumor cells.

Regression of AK7 malignant mesothelioma established in immunocompetent mice following intratumoral gene transfer of interferon gamma.

Cancer Gene Ther 2003 Jun;10(6):481-90 (ISSN: 0929-1903)

Cordier Kellerman L; Valeyrie L; Fernandez N; Opolon P; Sabourin JC; Maubec E; Le Roy P; Kane A; Legrand A; Abina MA; Descamps V; Haddada H

Institut Gustave Roussy, 94805 Villejuif Cedex, France.

Malignant mesothelioma (MM) is a lethal tumor linked with a prior exposure to asbestos in which limited progress has been made so far using conventional therapies. MM is an example of a "nonimmunogenic" tumor characterized by a fibrous stroma and an absence of infiltrating T lymphocytes. High levels of transforming growth factor-beta (TGF-beta) produced by mesothelioma cells have been related to the immune tolerance towards the tumor. In order to evaluate the effect of local delivery of cytokines such as interferon gamma (IFN-gamma) by gene transfer, we characterized and used a murine model, AK7, which appeared very similar to human mesothelioma. AK7 cells expressed low levels of major histocompatibility class I and class II antigens and secreted high levels of latent TGF-beta. The TGF-beta pathway in AK7 cells is operative but inefficient because endogenous TGF-beta is predominantly inactive. Treatment of pre-established AK7 tumors by direct intratumoral injection of an adenovirus vector expressing murine IFN-gamma, Ad.mIFN-gamma, led to significant tumor regression. Peripheral tumor infiltration by CD4+ and CD8+ T lymphocytes in the treated tumors appeared to be because of the induction of an immune response. Tumor relapse was observed, which could be due to local TGF-beta secretion by remaining tumor cells.

Analysis of the immunologic response generated by Ad.IFN-beta during successful intraperitoneal tumor gene therapy.

Mol Ther 2002 Aug;6(2):210-8 (ISSN: 1525-0016)

Odaka M; Wiewrodt R; De Long P; Tanaka T; Zhang Y; Kaiser L; Albelda S

Thoracic Oncology Research Laboratory, University of Pennsylvania Medical Center, Philadelphia, 19104, USA.

One promising therapeutic approach to intracavitary tumors, such as malignant mesothelioma and ovarian cancer, is immuno-gene therapy. In a previous study, intraperitoneal (i.p.) instillation of an adenoviral vector encoding the mouse interferon-beta gene (Ad.muIFN-beta) was shown to eradicate established mesothelioma tumors in the peritoneal cavity of immune competent, but not immunodeficient mice. The goal of this study was to understand more completely the kinetics and mechanisms of this immune-mediated response. Two days after a single intraperitoneal (i.p.) injection of Ad.muIFN-beta into BALB/c mice with established tumors, the response in the peritoneum was characterized by an influx of activated natural killer (NK) cells, polymorphonuclear (PMN) cells, and macrophages with minimal infiltration into the tumor nodules. However, depletion of PMN or NK cells after Ad.IFN-beta treatment had only minimal effects. At later time points (up to 10 days after Ad.IFN-beta i.p.), a large influx of CD4(+) and activated CD8(+) T cells was present in the peritoneal fluid and within the tumor nodules. The CD8(+) T cells had cytolytic activity, and adoptive transfer of peritoneal exudate cells (obtained by peritoneal lavage) resulted in effective tumor cell killing. Antitumor effects of Ad.IFN-beta may be different in different tumor types or in different anatomic locations. However, these results demonstrate that tumor-specific CD4(+) and CD8(+) T cells are the key effector cells for tumor eradication in this model

Intrapleural infusion of activated macrophages and gamma-interferon in malignant pleural mesothelioma: a phase II study.

Chest 2002 Jun;121(6):1921-7 (ISSN: 0012-3692)

Monnet I; Breau JL; Moro D; Lena H; Eymard JC; Menard O; Vuillez JP; Chokri M; Romet-Lemonne JL; Lopez M

Department of Pneumology, Center Hospitalier Intercommunal de Creteil, Creteil, France.

STUDY OBJECTIVES: Intrapleural immunotherapy has shown some activity in patients with malignant mesothelioma. We conducted a multicentric pilot phase II study to evaluate the tolerance and the activity of intrapleurally infused autologous human activated macrophages (AM Phi) in patients with stage IA, IB, and IIA malignant pleural mesothelioma (MPM).

DESIGN: AM Phi derived from in vitro monocyte culture were infused into the pleura of patients every week for 8 consecutive weeks. Each infusion was followed 3 days later by an intrapleural injection of 9 millions units of gamma-interferon (gamma-IFN) in an attempt to prolong the in vivo activation of infused AM Phi. Response was assessed by CT scan and thoracoscopy when possible. If the patient's disease progressed after AM Phi treatment, an additional treatment was left to the choice of the investigator.

PATIENTS: Nineteen patients with histologically proven stage IA, IB, or IIA MPM were enrolled. Two patients were excluded before any AM Phi infusion because of complications impeding infusion. Seventeen patients were actually treated. After completion of the AM Phi cellular therapy, 10 patients were treated with chemotherapy as their diseases progressed.

RESULTS: The overall response rate of patients actually treated was 14%. When including the two patients enrolled but not treated, the overall response "in intention to treat" was 11%; two patients had a partial response, with a duration of response of 30 months and 3 months, respectively. One patient, who could not be evaluated by thoracoscopy because of pleural symphysis, is still alive without any clinical or radiologic sign of disease 69 months after treatment. No major adverse effects were observed during the infusion of either AM Phi or gamma-IFN, and there was no interruption of treatment because of toxicity. However, symphysis was observed in 7 of 14 patients who received the complete treatment. The median survival of patients actually treated, including those who received chemotherapy after AM Phi, was 29.2 months.

CONCLUSION: Combined infusion of AM Phi and gamma-IFN was well tolerated in patients with MPM; however, it had limited antitumor activit

Combined regimen of cisplatin, doxorubicin, and alpha-2b interferon in the treatment of advanced malignant pleural mesothelioma: a Phase II multicenter trial of the Italian Group on Rare Tumors (GITR) and the Italian Lung Cancer Task Force (FONICAP).

Cancer 2001 Aug 1;92(3):650-6 (ISSN: 0008-543X)

Parra HS; Tixi L; Latteri F; Bretti S; Alloisio M; Gravina A; Lionetto R; Bruzzi P; Dani C; Rosso R; Cosso M; Balzarini L; Santoro A; Ardizzoni A

Department of Medical Oncology and Hematology, Istituto Clinico Humanitas, Via Manzoni 56, 20089 Rozzano-Milano, Italy. hector.sotoparra@humanitas.it.

BACKGROUND: The cisplatin-doxorubicin combination has shown moderate activity in malignant pleural mesothelioma (MPM; objective response, 25%), and preclinical studies suggest that interferons (IFNs) may have an antiproliferative effect on mesothelioma cell lines with a marked increase in cisplatin cytotoxicity. Therefore, the combined chemoimmunotherapy regimen is an worthwhile approach to evaluate in a Phase II trial.

METHODS: From December 1995 to June 1999, 37 previously untreated patients with MPM were treated with cisplatin 60 mg/m(2) intravenously on Day 1 plus doxorubicin 60 mg/m(2), recycled every 3-4 weeks and IFN-alpha-2b, 3 x 10((6)) international units subcutaneously 3 times a week for a total of 6 courses or until progression. Inclusion criteria were histologic diagnosis of MPM and measurable disease defined by computed tomography scan or magnetic resonance imaging.

RESULTS: Thirty-four patients were assessable for toxicity and 35 for efficacy according to World Health Organization criteria. One hundred thirty-five courses were administered with a median of 4 cycles per patients. Seventy-six percent of patient presented at least 1 episode of severe myelosuppression (Grade 3 and 4). Severe anemia and thrombocytopenia occurred in 30% and 24% of patients, respectively. Sixty percent of patients presented constitutional symptoms. In the 35 patients assessable for response, the overall response rate was 29% (95% confidence interval, 15-47%). The median duration of response was 8.4 months. With a median follow-up of 19.6 months, the median survival was 9.3 months. One- and 2-year survival was 45% and 34%, respectively.

CONCLUSIONS: This combined regimen has definite activity in MPM. However, toxicity, particularly myelosuppression and fatigue, is not negligible and may limit its application. [Copyright 2001 American Cancer Society.].

Control of cell cycle progression in human mesothelioma cells treated with gamma interferon.

Oncogene 2001 Mar 1;20(9):1085-93 (ISSN: 0950-9232)

Vivo C; Levy F; Pilatte Y; Fleury-Feith J; Chretien P; Monnet I; Kheuang L; Jaurand MC

INSERM E 99.09, Universite Paris Val de Marne Paris XII (EA 2345), Faculte de Medecine, 8 rue du General Sarrail, 94010, Creteil Cedex, France.

Recombinant human interferon gamma (r-hu-IFNgamma) exerts both antitumoral activity in the early stages of human malignant mesothelioma and a cytostatic effect in human mesothelioma (HM) cell lines in vitro. The antiproliferative effect of interferons (IFNs) reported in a variety of cells has been attributed to several mechanisms. In order to progress in the understanding of HM cell growth modulation by r-hu-IFNgamma, modifications of cell cycle progression and expression of key cell cycle regulator proteins in response to r-hu-IFNgamma were examined. Nine HM cell lines were studied, including one resistant to the antiproliferative effect of r-hu-IFNgamma. Except in the resistant cell line r-hu-IFNgamma produced an arrest in the G1 and G2-M phases of the cell cycle, associated with a reduction in both cyclin A and cyclin dependent kinase inhibitors (CDKIs) expression. Moreover cyclin B1/cdc2 activity was decreased. The present study provides the first evidence of a G2-arrest in r-hu-IFNgamma-treated HM cell lines and indicates that HM cell lines, despite their tumorigenic origin still support cell cycle control. The cell cycle arrest induced by r-hu-IFNgamma seems to depend on cyclin regulation through p21(WAF1/CIP1)- and p27(Kip1)-independent mechanisms and is not directly related to the induced DNA damage.