Research Report 2010 - MDC

For the delivery of naked DNA into cells or tissues agreat variety of procedures is employed in vitro and invivo. Among other physical delivery systems jet-injectionhas developed to an applicable gene transfer technology,which allows gene transfer of small amounts ofnaked DNA into different tissue types with deeper penetrationand improved dispersion.At the Clinic for Surgical Oncology, Charité, Berlin aphase I clinical trial (DeReGe 62) was conducted to evaluatesafety, feasibility and efficiency of intratumoral jetinjectiongene transfer of β-galactosidase (LacZ)-reporter expressing plasmid-DNA into skin metastasesfrom breast cancer and melanoma. Seventeen patientswere enrolled and treated with jet-injection into a singlecutaneous lesion. In the study the safety and efficiencyof the jet-injection gene transfer was shown. Thestudy revealed efficient LacZ mRNA- and proteinexpressionin all treated lesions and rapid clearance ofplasmid-DNA from patient’s blood. The treatment waswell tolerated by all patients and no side effects wereexperienced, indicating clinical applicability of this nonviralapproach for local tumor gene therapy.Use of the minimalistic MIDGE vector forimproved safety, gene transfer and expression inclinical application of gene therapyW. Walther, S. Burock, D. Kobelt, J. Aumann, U. Stein,P. M. Schlag. In cooperation with B. Wittig and M. Schmidt(MOLOGEN AG, Berlin), U. Trefzer (Charité) and I. FichtnerIn result of our clinical gene transfer trial we currentlymake great efforts for further optimization of the nonviralvector by removing unnecessary sequences for theimprovement of transfer- and expression efficiency. Theminimalistic immunologically defined gene expression(MIDGE, Mologen AG, Berlin) vector system provides anonviral linear DNA molecule that is depleted of anybacterial origin, antibiotic resistance sequences or replicationbackbone sequences. This makes MIDGE of highvalue with respect to regulatory requirements for productsafety in clinical applications. Thus, MIDGE vectorsare substantially reduced in size, resulting in highertransfer efficiencies and improved transgene expression.Our in vitro analyses in different human tumor celllines revealed pronounced increase in transfer efficiencyand in transgene expression of the MIDGE vector systemcompared to plasmid-based vectors. The use of thissystem for expression of therapeutic genes, such ashuman TNF-α, mediated high-level expression in vitroand more importantly in vivo and resulted in effectiveantitumoral effects. These expression characteristicsmake MIDGE a good candidate for use in clinical genetherapy applications.Based on our pre-clinical and clinical experiences onnonviral gene transfer a new phase I trial will be initiatedto evaluate safety and efficiency of nonviral jet-injectionapplication of the TNF-α expressing MIDGE-basedvector in patients with skin metastasis from mela -noma. This dose-escalation study will reveal, whetherthe MIDGE-vector is efficiently expressing TNF-α afterthe intratumoral application by jet-injection. It will beimportant to correlate the applied vector dose and theamount of TNF-α expressed in the tumor lesion. Thesafety of the expressed TNF-α and the vector-DNA doseapplied will be examined and will be tested in a clinicalphase II trial, to evaluate the safety and efficacy ofthis approach for the combined modality treatment ofpatients with malignant melanoma.Selected PublicationsStein, U., Walther, W., Arlt, F., Schwabe, H., Smith, J., Fichtner, I., Birchmeier,W., Schlag, P.M. (2009) MACC1, a newly identified key regulator of HGF-MET signaling, predicts colon cancer metastasis. Nat Med. 15, 59-67Fritzmann J, Morkel M, Besser D, Budczies J, Kosel F, Brembeck FH, Stein U,Fichtner I, Schlag PM, Birchmeier W. (2009) A colorectal cancer expressionprofile that includes transforming growth factor beta inhibitor BAMBIpredicts metastatic potential. Gastroenterology 137; 165-175Kemmner W, Wan K, Rüttinger S, Ebert B, Macdonald R, Klamm U, MoestaKT (2008) Silencing of human ferrochelatase causes abundant protoporphyrin-IXaccumulation in colon cancer – a new tool for molecular imaging.FASEB J 22, 500-509Walther W, Siegel R, Kobelt D, Knösel T, Dietel M, Bembenek A, Aumann J,Schleef M, Baier R, Stein U, Schlag PM. (2008) Novel jet-injection technologyfor nonviral intratumoral gene transfer in patients with melanomaand breast cancer. Clin Cancer Res 14, 7545-53.Stein, U., Arlt, F., Walther, W, Smith, J., Waldman, T., Harris, E.D., Mertins,S.D., Heizmann, C.W., Allard, D., Birchmeier, W., Schlag, P.M., and Shoemaker,R.H (2006). The metastasis-associated gene S100A4 is a novel target ofβ-catenin/T-cell factor signalling in colon cancer. Gastroenterology 131,1486-1500.PatentsKemmner W, Schlag PMMicroarray für das Expression Profiling der kompletten zellulärenGlykosylierungsmaschineriePatent application: 9.10.2008Stein U, Lange C, Walther W, Schlag PM.Use of the human LRP/MVP promoter for a vector that can be induced bytherapy.Patent EP 1332219 10.5.2006US 7,297,535 20.11.2007Stein U, Schwabe H, Walther W, Schlag PM.Verwendung des neu-identifizierten Gens 7a5/Prognostin fürTumordiagnostik und Tumortherapie.7a5/Prognostin and its use for tumor diagnostics and therapy.Patent application US 10/564,823 2006Stein U, Walther W, Arlt F, Schlag PM.Methods for diagnosing metastasis by analysing mutations in β-catenin.Patent application US 60/847,547 2006Patent application EP 008333 2007Cancer Research 99