D. Kineman, Ph.D.
Lab Phone: 312-569-6983
Office and Laboratory Location:
Jesse Browh VA Medical Center
Research & Development
Bldg. 11A, Rm 6271, 6283, 6285
820 S. Damen Ave.
Chicago, IL 60612
University of Illinois at Chicago, Department of Medicine
Section of Endocrinology and Metabolism
1819 West Polk St., M/C 640
Chicago, IL 60612
BS - Michigan State University, East Lansing, MI
MS and Ph.D. - University of Georgia, Athens, GA
Postdoctoral Training - Medical University of South Carolina, Charleston, SC
Objective 1 - Growth hormone (GH) is a hormone synthesized in the anterior pituitary by a sub-population of cells called somatotropes. GH (as it's name implies) is vital for normal body growth during development but is also important in metabolic regulation in the adult. A primary focus of our laboratory is to understand how changes in the hormonal and metabolic environment alters GH synthesis and release particularly as it relates to changes in hypothalamic GH-releasing hormone, somatostatin and neuropeptide Y expression and synthesis of pituitary somatotrope receptors (GHRH-R, sst1-5 and GHS-R).
Objective 2 - The normal development, expansion and maintenance of the GH-producing pituitary cell population (somatotropes) is dependent on the hypothalamic neuropeptide, GH-releasing hormone (GHRH). GHRH acts to stimulate somatotrope proliferation, as well as GH synthesis and release, by activation of the cAMP intracellular signal transduction pathway. Chronic stimulation of the cAMP signaling pathway leads to somatotrope hyperplasia and GH-secreting pituitary tumors in both mice and humans. Somatostatin, also a hypothalamic neuropeptide, opposes the actions of GHRH and blocks cAMP accumulation and in the clinical setting can prohibit the growth of some pituitary tumors. We are currently using animal models to understand the mechanism by which cAMP causes pituitary cells to form tumors and how somatostatin inhibits pituitary tumor growth.
Development of a mouse model (rGHp,Cre) to selectively knockout genes on in the GH-producing cells of the pituitary
My laboratory has recently developed genetically-engineered mice that will allow for the inactivation of genes only in the pituitary somatotrope population. Using this model we can directly test the importance of various genes in the somatotrope population, without disturbing gene function in the rest of the body. This strategy takes advantage of a prokaryotic system in which the bacteriophage enzyme, Cre recombinase (Cre), can excise bacterial DNA which is flanked by specific palindromic sequences referred to as LoxP sites. This system has been extensively used to genetically engineer eukaryotic cell lines and mice to confer time- and tissue-specific inactivation of genes. In mice, the Cre transgene is expressed in the tissue of interest by use of a tissue-specific promoter. These mice are then crossbred to genetically engineered mice harboring a gene that has been modified by the insertion of two LoxP sites that flank critical functional elements of the gene to be disrupted (ie. the gene is "floxed"). In the absence of Cre the function of the floxed gene is unperturbed, however in the tissues where Cre is expressed the floxed DNA is excised and the gene is inactivated. To specifically study the somatotrope population we have generated a mouse that expresses the Cre in the GH-producing cells of the anterior pituitary using the rat GH gene promoter (rGHp). To test if Cre is expressed in the pituitary and is functional we crossbred the rGHp,Cre mice to the reporter mice, Z/AP, generated by Dr. Corinne Lobe (Sunnybrook & Women's College Health Sciences Centre, Totonto, Canada). In tissues where active Cre is present it excises floxed DNA of the reporter Z/AP gene resulting in the expression of human alkaline phosphatase which in the presence of substrate turns the tissue blue. Below is a figure demonstrating Cre activity in the anterior pituitary gland of rGHp,Cre X Z/AP mice.
Raul Luque, PhD Research Assistant
(from University of Cordoba, Spain)
Selected Recent Publications and Manuscripts in Review:
Park S, Peng X-D, Frohman LA, Kineman RD. Expression analysis of hypothalamic and pituitary components of the growth hormone-axis in fasted and streptozotocin-treated NPY-intact (NPY+/+) and NPY-knockout (NPY-/-) mice. Neuroendocrinology 81(6):360-371, 2005
Kim E, Sohn S, Lee M, Jung J, Kineman RD, Park S. Differential responses of the growth hormone axis in two rat models of streptozotocin-induced insulinopenic diabetes. Journal of Endocrinology 188(2):263-270, 2006
Luque RM, Gahete M, Kineman RD Examination of the direct effects of metabolic factors on somatotrope function in a non-human primate model (Papio anubis). J. Molecular Endocrinology 37:1-15, 2006
Luque RM, Kineman RD. Impact of obesity on the growth hormone (GH)-axis: evidence for a direct inhibitory effect of hyperinsulinemia on pituitary function. Endocrinology 147:2754-2763, 2006
Luque RM, Kineman RD, Duran M, Gracia-Navarro S, Gracia-Navarro F, Malagon MM, Castano JP Identification of the somatostatin receptor subtypes that control growth hormone secretion from pig somatotropes. Endocrinology 147(6):2902-2908, 2006
Luque RM, Gahete M, Hochgeschwender U, Kineman RD. Evidence that endogenous somatostatin (SST) inhibits adrenocorticotropin (ACTH) and ghrelin expression by independent pathways. American Journal of Physiology; Endocrinology and Metabolism March 291:E395-403, 2006
Sandoval R, Xue J, Tian X, Barrett K, Pilkinton M, Baida G, Ucker DS, Raychaudhuri P, Kineman RD, Luque RM, Baida G, Zou X, Valli, VE, Cook JL, Kiyokawa H, Colamonici OR. A mutant allele of BARA/LIN-9 rescues the cdk4-/- phenotype by releasing the repression on E2F-regulated genes. Experimental Cell Research 312:2465-75, 2006
Luque RM, Peinado JR, Gracia-Navarro F, Broglio F, Ghigo E, Kineman RD, Malagon MM, Castano JP. Cortistatin mimics somatostatin by inducing a dual, dose-dependent stimulatory and inhibitory effect on growth hormone secretion in somatotropes. Journal of Molecular Endocrinology 36(3):547-56, 2006
Luque RM, Park S, Kineman RD . Severity of the catabolic condition in response to fasting differentially modulates hypothalamic expression of growth hormone-releasing hormone (GHRH) in the mouse; potential role for corticotropin releasing hormone (CRH). Accepted pending revisions Endocrinology (June 2006)
Luque RM, Huang Z, Mazzone T, Kineman RD Leptin replacement enhances growth hormone (GH)-axis function and circulating ghrelin levels in ob/ob mice, independent of food intake and weight loss. Accepted pending minor revisions American Journal of Physiology Endocrinology and Metabolism (June 2006)
Taboada GF, Luque RM, Bastos W, Guimar„es RFC, Marcondes JB, Chimelli LMC, Mata PJP, Niemeyer Filho P, Carvalho DP, Kineman RD, Gadelha MR Somatostatin receptor subtype quantitative gene expression in human somatotropinomas and non-functioning pituitary adenomas Accepted pending minor revisions European Journal of Endocrinology (June 2006)
in Progress - Presentations for Summer 2004/2005 Scientific Meetings:
Luque RM, Kineman RD Studies examining the hypothalamic and pituitary effect of leptin replacement on growth hormone (GH)- and adrenal-axis function, as well as ghrelin release and synthesis in the ob/ob mouse. 6th International Congress of Neuroendocrinology, Pittsburg, PA June 2006
Luque RM, Kineman RD Duration of fasting differentially modulates the hypothalamic-pituitary-growth hormone (GH) axis of the mouse 88th Annual Meeting of the Endocrine Society, Boston June 2006
Kineman RD, Gahete MD, Luques RM Identification of a mouse ghrelin variant that contains Exon 2, Intron 2 and Exon 3, but lacks Exon 4 and Exon 5; evidence for tissue specific regulation 88th Annual Meeting of the Endocrine Society, Boston June 2006
Wang Z, Kineman RD, Luque RM, Shirai T, Lantvit DD, Unterman TG, Prins GS, Swanson SM Growth hormone receptor mRNA is upregulated, while IGF-I receptor mRNA is downregulated during prostate carcinogenesis in the probasin/SV40 TAg rat 88th Annual Meeting of the Endocrine Society, Boston June 2006