Abstract
The prognosis of pancreatic adenocarcinoma is poor and current treatment is for the most part ineffective. We describe here a novel treatment strategy using a mouse model system for pancreatic cancer. Human embryonic epithelial cells have been genetically modified to express the cytochrome P450 2B1 enzyme under the control of a CMV immediate-early promoter. This CYP2B1 gene converts oxazaphosphorines (ifosfamide or cyclophosphamide) to their active cytotoxic compounds, phosphoramide mustard, which alkylates DNA, and acrolein, which alkylates proteins. A number of assays were performed to demonstrate the CYP2B1 gene function as well as toxic effects on neighbouring cells (bystander effect). The cells were then encapsulated in a cellulose sulphate formulation shown to be well tolerated in the pancreas of immunocompetent mice, and injected 1 cm away from pre-established tumours derived from a human pancreatic tumour cell line (PaCa-44). Intraperitoneal administration of low-dose ifosfamide to tumour bearing mice that received the encapsulated cells results in partial or even complete tumour ablation. Such an in situ chemotherapy strategy utilizing genetically modified cells in an immuno-protected environment may prove useful for solid tumour therapy in man.
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References
Ahlgren JD (1996): Chemotherapy for pancreatic carcinoma. Cancer; 78: 654–663.
Brock N, Hilgard P, Peukert M, Pohl J and Sindermann H (1988): Basis and New developments in the Field of Oxazaphosphorines. Cancer Investigation; 6: 513–532.
Carmichael J, Fink U, Russell RCG, Spittle MF, Harris AL, Spiessi G, Blatter J (1996): Phase II study of gemci-tabine in patients with advanced pancreatic cancer. Br J Cancer; 73: 101–105.
Chen L and Waxman J (1995): Intratumoral activation and enhanced chemotherapeutic effect of oxazaphosphorines following cytochrome P-450 gene transfer: development of a combined chemotherapy/cancer gene therapy strategy. Cancer Research; 55: 581–589.
Crandell RA, Fabricant CG, Nelson-Rees WA (1973): Development, characterization, and viral susceptibility of a feline (Felis catus) renal cell line (CRFK). In Vitro; 9: 176–185.
Connors TA (1995): The choice of prodrugs for gene directed enzyme prodrug therapy of cancer. Gene Therapy; 2: 702–709.
Cosset FL, Takeuchi Y, Battini JL, Weiss RA and Collins MK (1995): High-titer packaging cells producing recombinant retroviruses resistant to human serum. J Virol; 69: 7430–7436.
Dirven HAAM, van Ommen B, van Blaederen PJ (1996): Glutathione conjugation of alkylating cytostatic drugs with a nitrogen mustard group and the role of glutathione S-transferases. Chem Res Toxicol; 9: 351–360.
Donato MT, Gomez-Lechon MJ, Castell JV (1993): A microassay for measuring cytochrome P450IA1 and P450IIB1 activities in intact human rat hepatocytes cultured on 96-well plates. Anal Biochem; 213: 29–33.
Günzburg WH, Karle P, Mrochen S, Sparmann G, Sailer R, Klein D, Uckert W and Salmons B (1997): Regulated gene expression after retroviral vector-mediated delivery of cancer-relevant therapeutic genes. Recent Results Cancer Res; 144: 116–126.
Kalthoff H, Löhr M, Roeder C, Schmiegel W (1995): Das Pankreaskarzinom: Zellbiologie, Matrixproteine und Wachstumsregulation. In: Erkrankungen des exkretorischen Pankreas; Adler G, Fölsch UR, Mössner J, Singer MV. eds. Fischer, Jena: 2nd edition, p.385–404.
Kedzie KM, Balfour CA, Escobar GY, Grimm SW, He YA, Pepperl DJ, Regan JW, Stevens JC, Halpert JR (1991): Molecular basis for a functionally unique cytochrome P450IIB1 variant. J Biol Chem; 266: 22515–22521.
Keizer HJ, Ouwerkerk J, Welvaart K, van der Velde CJH, Cleton FJ (1995): Ifosfamide treatment as a 10-day continuous intravenous infusion. J Cancer Res Clin Oncol; 121: 297–302.
Kurowski V, Wagner T (1993): Comparative pharmacokinetics of ifosfamide, 4-hydroxyifosfamide, chloracetaldehyde, and 2- and 3-dechloroethylifosfamide in patients on fractionated intravenous ifosfamide therapy. Cancer Chemother Pharmacol; 33: 36–42.
Löhr M, Trautmann B, Göttler M, Peters S, Zauner I, Maillet B, Klöppel G (1994): Human ductal adenocarcinomas of the pancreas express extracellular matrix proteins. Br J Cancer; 69: 144–151.
Löhr M, Trautmann B, Peters S, Zauner I, Liebe S, Klöppel G, Kreuser ED (1995): Expression and function of integrins in human pancreatic adenocarcinoma. Pancreas; 12: 248–259.
Löhr M 1996: Zell- und Molekularbiologie der Bindegewebsreaktion bei Pankreatitis und Pankreaskarzinom. Solvay, Hannover.
Löhr M, Müller P, Karle P, Stange J, Mitzner S, Nizze H, Liebe S, Salmons B, Günzburg WH (1997): Targeted chemotherapy by encapsulating cells engineered to deliver CYP2B1, an ifosfamide activating cytochrome P450 gene. Gene Therapy; submitted
McClane SJ, Hamilton TE, DeMatteo RP, Burke C and Raper SE(1997): Effect of adenoviral early genes and the host immune system on in vivo pancreatic gene transfer in the mouse. Pancreas; 75: 236–245.
Pear WS, Nolan GP, Scott ML, Baltimore D (1993): Production of high-titer helper-free retroviruses by transient transfection. Proc Natl Acad Sci (USA); 90: 8392–8396.
Reyes G, Villanueva A, Garciá C, Sancho FJ, Piulats J, Lluis F, Capellâ G (1996): Orthotopic xenografts of human pancreatic carcinomas acquire genetic aberrations during dissemination in nude mice. Cancer Res; 56: 5713–5719.
Rosewicz S, Wiedenmann B (1997): Pancreatic carcinoma. Lancet; 349: 485–489.
Sailer RM, Stange J, Mitzner S, Heinzmann U, Salmons B, Günzburg WH (1997): Encapsulated cells producing retroviral vectors for in vivo gene therapy. Submitted.
Stange J, Mitzner S, Dautzenberg H, Ramlow W, Knippel M, Steiner M, Ernst B, Schmidt R, Klinkmann H (1993): Prolonged biochemical and morphological stability of encapsulated liver cells—a new method. Biomat Art Cells & Immob Biotech; 21: 343–352.
Takeuchi Y, Porter CD, Strahan KM, Preece AF, Gustafsson K, Cosset FL, Weiss RA, Collins MK (1996): Sensitization of cells and retroviruses to human serum by (alpha 1–3) galactosyl transferase. Nature; 379: 85–88.
WHO (1979): WHO handbook for reporting results of cancer treatment. WHO, Geneva.
Wright J, Tretyakov O, Ayash L, Elias A, Rosowsky A and Frei EI (1995): Analysis of 4-hydroxycyclophos-phamide in human blood. Analyt Biochem; 224: 154–158.
Zheng JJ, Chan KK, Muggia F (1994): Preclinical pharmacokinetics and stability of isophosphoramide mustard. Cancer Chemother Pharmacol; 33: 391–398.
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Karle, P. et al. (1998). Intratumoral Injection of Encapsulated Cells Producing an Oxazaphosphorine Activating Cytochrome P450 for Targeted Chemotherapy. In: Walden, P., Trefzer, U., Sterry, W., Farzaneh, F., Zambon, P. (eds) Gene Therapy of Cancer. Advances in Experimental Medicine and Biology, vol 451. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-5357-1_16
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DOI: https://doi.org/10.1007/978-1-4615-5357-1_16
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