Development of the limbs and external genitalia require the coordination of many different regulatory pathways and signaling events. While it is now clear that important components of the limb and genital appendage gene networks are shared, remarkably little is known about the cis- regulatory elements that contribute the transcriptional control of these networks. The Tbx4, Pitx1, and Isl1 transcription factors are among a very small number of hind limb-specific genes that function in the hind limb but not the forelimb. Though work in animal models and humans has shown a central role for these factors in leg formation, we have almost no knowledge of the functional interactions these transcription factors make with the network of cis-regulatory elements that promote limb development. In addition, our preliminary data indicates that at least of one these factors, Tbx4, is also an important regulator of genital growth, suggesting that these genes may exhibit overlapping functional interactions with appendage cis-regulatory elements active in the hind limbs and genitals. The objective of this proposal is to determine how Pitx1, Tbx4, and Isl1 influence the development of appendages, by investigating their interactions with the suite of cis- regulatory elements active in the developing hind limbs and genitalia. This objective will be accomplished by 1) identifying the set of enhancer elements that are bound and directly regulated by Pitx1 in the hind limbs and genitals, 2) determining whether Tbx4 directs the outgrowth of the legs and external genitalia via similar mechanisms, and 3) establishing whether the initiation of hind limb bud and genital bud formation is regulated by Isl1 through a common set of regulatory interactions. The knowledge gained from these studies will help to achieve our long-term goal of understanding how the disruption of specific regulatory interactions can lead to malformations of the limbs and genitals in humans.

Public Health Relevance

The Tbx4, Pitx1, and Isl1 genes are essential for controlling formation of the leg, and mutations in two of these genes have been implicated in severe lower limb malformations in humans. The goal of this study is to understand how these genes work together to promote proper development of the leg and to determine what role they play in formation of the external genitalia. This study will improve our understanding of how mutations result in congenital birth defects of the limbs and genitals.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Project (R01)
Project #
5R01HD081034-05
Application #
9494644
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Coulombe, James N
Project Start
2014-08-15
Project End
2019-04-30
Budget Start
2018-05-01
Budget End
2019-04-30
Support Year
5
Fiscal Year
2018
Total Cost
Indirect Cost
Name
University of Georgia
Department
Genetics
Type
Schools of Arts and Sciences
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602
Ching, Saunders T; Infante, Carlos R; Du, Wen et al. (2018) Isl1 mediates mesenchymal expansion in the developing external genitalia via regulation of Bmp4, Fgf10 and Wnt5a. Hum Mol Genet 27:107-119
Infante, Carlos R; Rasys, Ashley M; Menke, Douglas B (2018) Appendages and gene regulatory networks: Lessons from the limbless. Genesis 56:
Wang, Jialiang S; Infante, Carlos R; Park, Sungdae et al. (2018) PITX1 promotes chondrogenesis and myogenesis in mouse hindlimbs through conserved regulatory targets. Dev Biol 434:186-195
Thompson, Abbey C; Capellini, Terence D; Guenther, Catherine A et al. (2018) A novel enhancer near the Pitx1 gene influences development and evolution of pelvic appendages in vertebrates. Elife 7:
Infante, Carlos R; Mihala, Alexandra G; Park, Sungdae et al. (2015) Shared Enhancer Activity in the Limbs and Phallus and Functional Divergence of a Limb-Genital cis-Regulatory Element in Snakes. Dev Cell 35:107-19