Homeobox genes in cardiovascular development

As summarized earlier, a surprisingly large number of different homeobox genes are expressed in the developing heart. Some are clearly important, as demonstrated by mouse gene ablation studies. For example, knockout of Nkx2-5 or Hoxa-3 function is embryonic lethal due to defects in cardiovascular development. However, gene ablation studies indicate that other homeobox genes that show cardiovascular expression are either not required for heart development or their function is effectively complemented by a redundant gene activity. Given the number of closely related homeobox genes that are expressed in the heart (and the rate at which new genes are being discovered), this is very likely to be the case for at least some homeobox gene activities. At present little is known of the precise mechanism of action of homeobox genes in embryonic development. This statement applies to homeobox genes in general, not just to genes involved in cardiovascular development. There is a popular view that homeobox genes are master regulators that control expression of a large number of downstream genes. In at least some cases, e.g., the eyeless gene of Drosophila (Holder et al., 1995), homeobox genes appear to be capable of activating and maintaining a very complex developmental program. Significantly, the eyeless gene is able to initiate eye development at numerous ectopic locations. Increasing evidence, however, suggests that genes of this type may be rather rare. Certainly there is no evidence to date that any of the homeobox genes expressed in the heart are able to initiate the complete heart development pathway. This is probably best understood in the case of the tinman gene in Drosophila, which, although absolutely required for heart development, is not capable of initiating the cardiac development pathway in ectopic locations (Bodmer, 1993). This conclusion is supported by studies of the vertebrate tinman-related gene Nkx2-5. Gene ablation studies show that Nkx2-5 is essential for correct cardiac development (Lyons et al., 1995) but is not able to initiate the regulatory pathway leading to cardiac development when expressed ectopically (Cleaver et al., 1996; Chen and Fishman, 1996). If most homeodomain proteins are not direct regulators of a differentiation pathway, what is their role during organogenesis? The cardiovascular homeobox gene about which most is known at the mechanistic level is gax (Smith et al., 1997). A number of experiments indicate that the Gax protein is involved in the regulation of cell proliferation and that it interacts with components of the cell cycle regulation machinery. Indeed, over recent years, the idea that at least some homeobox genes play their role in organogenesis through regulation of proliferation has been developed in some detail by Duboule (1995). Further evidence that this mechanism of homeobox activity is important, especially during organogenesis, comes from studies of the Hox11 homeobox gene, which is absolutely required for development of the spleen in mouse (Roberts et al., 1994). Studies indicate that Hox11 is able to interact with at least two different protein phosphatases, PP2A and PP1, which in turn, are involved in cell cycle regulation (Kawabe et al., 1997). It is quite clear that research in future years will need to focus on the precise mode of action of the different homeodomain proteins if we are to understand their role in the development of the cardiovascular system.