Winter, Desmond C. M.D., F.R.C.S.I.1; Shields, Conor J. M.D., A.F.R.C.S.I.1; Kirwan, William O. M.Ch., F.R.C.S.I.1 Author Information

PURPOSE: Duodenal adenomas arise in more than 90 percent of patients with familial adenomatous polyposis (FAP). Management of severe duodenal disease remains difficult and controversial. This study investigates transfer of functional wild-type adenomatous polyposis coli (APC) gene under conditions of varying pH and bile concentrations into a somatic duodenal cancer cell line (HUTU-80) as a prelude toin vivogene therapy in the management of severe duodenal disease. METHODS: In vitrotransfection of human APC gene was performed on a human duodenal adenocarcinoma cell line (HUTU-80) by use of a liposomal vector. Different concentrations (5 percent and 10 percent) of human bile and varying pH (6 percent and 8 percent) were used during APC gene transfer to assess their effects on transfection efficiency. The effect of bile on the proliferation of HUTU-80 cells was evaluated by means of a colorimetric chemosensitivity assay with sulforhodamine B (SRB). The duration of APC transgene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Exogenous APC transgene was expressed in HUTU-80 cell line for 6 days after transfection. Differing pH did not affect APC gene transfer into the duodenal cell line with similar transgene expression to controls, but APC transfection efficiency was reduced semiquantitatively in the presence of bile. Coculturing with human bile (5 percent and 10 percent) did not affect the proliferation of HUTU-80. CONCLUSION: This study demonstrates transfer of APC gene into a duodenal epithelial cell line with prolonged transgene expression. Liposome-mediated APC gene transfer to the duodenum is feasible even in the presence of bile and varying pH, raising the potential of future gene therapy for this extremely difficult to treat condition.

PURPOSE: Familial adenomatous polyposis is a highly penetrant, autosomal dominant disease resulting from a germline mutation of the adenomatous polyposis coli gene. Besides colorectal polyps and cancer, more than 90 percent of familial adenomatous polyposis patients also develop duodenal polyposis with an approximately 5 percent lifetime risk of malignant transformation. Because adenomatous polyposis coli protein has a "gatekeeper role" in the adenoma-carcinoma sequence, replacing its function may reduce polyp formation. We studied the functional outcome of peroral, liposome-mediated adenomatous polyposis coli gene replacement therapy in a multiple intestinal neoplasia mouse model. METHODS: Twenty multiple intestinal neoplasia mice, heterozygous for the human homologue adenomatous polyposis coli gene, were randomly assigned to three groups: no treatment (n = 8); control plasmid containing green fluorescence protein reporter gene (n = 6); and plasmid containing the full-length adenomatous polyposis coli gene (n = 6). For the adenomatous polyposis coli-treated and green fluorescence protein reporter gene-treated groups, each mouse received the appropriate plasmid complexed with liposome, administered twice per week by oral gavage regime. Treatment lasted four weeks and all animals were killed at the end of treatment period with harvesting of intestinal tissue for polyp number estimation. RESULTS: There was a statistically significant 25 percent reduction in the total number of polyps in the adenomatous polyposis coli-treated (73.1 +/- 1.4) group compared with untreated control (97.8 +/- 5.3, P < 0.01, Tukey test) and multiple intestinal neoplasia mice treated with control green fluorescence protein gene (103.3 +/- 1.7, P < 0.01, Tukey test). CONCLUSION: Adenomatous polyposis coli gene dysfunction underlies tumorigenesis in familial adenomatous polyposis patients and multiple intestinal neoplasia mice. This in vivo study provides evidence to support a novel anti-adenoma strategy using enteral adenomatous polyposis coli gene replacement therapy.

yrs).Among 22 adenomas individually examined from 21 patients, 20 (91%) were accompanied by contiguous colonic metaplasia and 2 (9%) were not.Neither age nor gender was assoc=ated with the prevalence of colonic metaplasia or of adenomas.Ileal inflammation was noted in 27 patients overall (60%): 41% with Brooke ileostomy, 71% with ileal pouches.Ileal inflammation was not associated with the presence of either adenomas or colonic metaplasia.CONCLU-SIONS: Based on these data, ileal adenomas commonly develop in FAP patients after proctocolectomy, especially in those with ileal pouches.Most ileal neoplasms appear to arise in the setting of colonic metaplasia.