Diffuse leiomyomatosis associated with X-linked Alport syndrome: extracellular matrix study using immunohistochemistry and in situ hybridization.

Inherited diffuse esophageal leiomyomatosis, a benign tumor involving smooth muscle cells of the whole esophagus, is frequently associated with X-linked Alport syndrome, a hereditary disease of type IV collagen. Families with this condition are consistently found to have deletions encompassing the 5' ends of both the α5 chain of type IV collagen (COL4A5) and the α6 chain of type IV collagen (COL4A6) genes, always limited in COL4A6 to exons 1', 1, and 2. On the contrary, patients with COL4A5/COL4A6 deletions extending further into COL4A6 display no such tumors. Despite the deletion, a COL4A6 transcript including exon 4, but not exon 3, was found in a tumor sample, raising the possibility of the involvement of a truncated α6(IV) chain in the tumorous process. Using immunohistochemistry and in situ hybridization methods, we analyzed the expression and distribution of the α6 chain of type IV collagen in tumors in comparison with that of normal, fetal, and mature esophagus. We also studied associated changes in tumor basement membrane composition and in tumor-cell integrin subunit distribution. No labeling with α6(IV) antibodies was detected in tumors, ruling out the hypothesis of a stably integrated truncated α6(IV) chain in tumor basement membranes. In contrast, despite the deletions of the first two exons of the gene and its 5' end, a COL4A6 transcript is clearly expressed by tumor cells. This finding raises the question of a potential role for this RNA in the tumor process. The absence of the α6(IV) chain is associated with the absence of the α5(IV) chain, as was suggested by the COL4A5 deletion. An additional striking feature is the absence of the βl chain of laminin in tumor basement membranes and the lack of or uneven expression of the α5 integrin subunit. These findings show that dramatic changes in the composition of the matrix and the expression of integrin receptors also occur in this benign tumorous process.