Director, Pediatric Surgical Research Laboratories, Massachusetts
General Hospital
Marshall K. Bartlett Professor of Surgery, Harvard Medical School
185 Cambridge Street, CPZN 6206
Boston, MA 02114
617-724-1600 p
617-726-5057 f
pdonahoe@partners.org
Summary of Research
Dr. Donahoe's research program has been centered upon understanding the molecular mechanisms of growth in utero, with the hypothesis that negative growth regulators in the fetus could be developed as regulators of tumor growth. She focused on Mullerian Inhibiting Substance (MIS) which is produced by the normal male fetal testis and causes the female reproductive tracts consisting of the uterus, Fallopian tubes, and upper vagina, to regress in the normal male embryo. She and her colleagues purified MIS, cloned its gene and that of its receptors, studied its regulation and the genes that MIS in turn regulates, and examined the effects of MIS against female and male reproductive tumors for clinical application of MIS as an anticancer agent. Using an assay to detect Mullerian Inhibiting Substance developed by David MacLaughlin, she and her colleagues have established normative MIS levels for children of various ages and used this subsequently to study children with intersex abnormalities and undescended testes, undervirilization boys, and overvirilization girls, as well as patients with granulosa cell tumors.
The laboratory receives most of its funding from the National Institute of Health (87%) with 13% from other sources including foundations and philanthropy. The work for MIS has been continuously funded by the National Cancer Institute and by the National Institute of Child Health and Development for 30 years.
Dr. Donahoe's group identified tumors of Mullerian duct origin as targets for MIS and found that ovarian, and most recently, endometrial and cervical cancers express MIS type II receptors and are growth inhibited by MIS. Given these findings, they are partnering with biotechnology firms to scale up production of MIS for use in preclinical and clinical trials, initially evaluating its use in ovarian cancer patients. Antibodies have been developed to the MIS type II receptor protein and vaccines against the MIS type II receptor are being made to target tumors of Mullerian duct origin.
Dr. Donahoe, who is the Director of the Germ Cell/Reproductive Tract Stem Cell Institutive for the Harvard Stem Cell Institute, is studying the role of stem cells in the normal repair that occurs after cyclic ovulation in the ovary. They have discovered a side population (SP) after flow cytometry, in cell lines derived from normal ovarian surface epithelium and find that this SP has characteristics and markers of stem cells. A similar SP has bee found in ovarian surface epithelial cancers emanating from MISRII T antigen transgenetic mice. This SP continues to respond to MIS after serial enrichment. We are working on the hypothesis that ovarian cancers occur due to abnormal function of the stem cells in the surface epithelium had are normally controlled by MIS.
After Dr. Shyamala Maheswaran elucidated p16 as an important component in the downstream signal transduction pathway of MIS in inhibition of tumor growth, Drs. Donahoe and MacLaughlin have been investigating combination therapies with MIS, employing therapeutics known to upregulate p16 and which prevent breakdown of p16.
The laboratory has also studied factors important in testis differentiation in coordination with the care of children with ambiguous genetalia. Dr. Nelson Arango and Dr. Donahoe are currently examining differentially expressed factors which may be important and specific for ovarian differentiation.
Another important focus of the laboratory is to understand the genetics of common congenital anomalies that require surgical correction. Dr. Donahoe is the Principal Investigator for a large Program Project funded by the NICHD to understand potential genetic defects leading to congenital diaphragmatic hernia (CDH) and the lethality caused by lung hypoplasia and pulmonary hypertension. Using a candidate gene approach in multiple animal models, including drosphila, chick, and rodents, candidate genes are being identified and studied in these animal model systems for complementation or improvement of lung hypoplasia. Cell lines have been established from patients with these defects and their families for interrogation of abnormalities in these candidate genes, comparing affected children to their parents and their sibs. The laboratory is also examining families with multiple affecteds with congenital diaphragmatic hernias. This infrastructure will be tested by mutational analyses of a number of candidate genes. Novel cytogenetic techniques such as array-based Comparative Genomic Hybridization and FISH are being used to detect microdeletions and microduplications, and to determine genes of interest in these regions. They are also employing genome-wide SNP analysis to detect loss of heterozygosity in consanguineous families with multiple affected patients with CDH.
The template established for congenital diaphragmatic hernia will be used to study patients, cared for by Dr. Donahoe and her clinical colleagues, and their families with common congenital anomalies of the urinary tract and of the gastrointestinal tract. It is the goal of these studies to identify genes involved in these defects and to design strategies that can be used, after detection of these defects in utero, to ameliorate the severity of the defects before the babies are born.
Representative Publications
Donahoe PK, Ito Y, Marfatia S, Hendren WH. A graded organ culture assay for the detection of Müllerian Inhibiting Substance. J Surg Res. 1977; 23:141-8.
Donahoe PK, Swann DA, Hayashi A, Sullivan MD. Müllerian duct regression in the embryo is correlated with cytotoxic activity against a human ovarian cancer. Science. 1979; 205:913-5
Donahoe PK, Fuller AF Jr, Scully RE, Guy SR, Budzik GP. Müllerian Inhibiting Substance inhibits growth of a human ovarian cancer in nude mice. Ann Surg. 1981; 194:472-80.
Trelstad RL, Hayashi A, Hayashi K, Donahoe PK. The epithelial-mesenchymal interface of the male rat Müllerian duct: Loss of basement membrane integrity and ductal regression. Develop Biol. 1982; 92:27-40.
Budzik GP; Powell SM; Kamagata S; Donahoe PK. Mullerian inhibiting substance fractionation by dye affinity chromatography. Cell 1983 Aug;34(1):307-14.
Donahoe PK, Hendren WH III. Perineal reconstruction in ambiguous genitalia infants raised as females. Annals of Surg. 1984; 200:363-71.
Fuller AF, Jr, Krane IM, Budzik GP, Donahoe PK. Müllerian Inhibiting Substance reduction of colony growth of human gynecologic cancers in a stem cell assay. Gynecol. Oncology, 1985; 22(2):135-48.
Cate RL, Mattaliano RJ, Hession C, Tizard R, Farber NM, Cheung A, Ninfa EG, Frey AZ, Gash DJ, Chow EP, Fisher RA, Bertonis JM, Torres G, Wallner BP, Ramachandran KL, Ragin RC, Manganaro TF, MacLaughlin DT, Donahoe PK. Isolation of the bovine and human genes for Müllerian Inhibiting Substance and expression of the human gene in animal cells. Cell. 1986; 45:685-98.
Pepinsky RB, Sinclair LK, Chow EP, Mattaliano RJ, Manganaro TF, Donahoe PK, Cate RL. Proteolytic processing of Müllerian Inhibiting Substance produces a transforming growth factor-B-like fragment. J. Biol. Chem. 1988; 263:18961-4.
Hudson PL, Dougas I, Donahoe PK, Cate RL, Epstein J, Pepinsky RB, MacLaughlin DT. An immunoassay to detect human Müllerian Inhibiting Substance in males and females during normal development. Journal of Clinical Endocrinology and Metabolism. 1990; 70(1):16-22.
Ragin RC, Donahoe PK, Kenneally MK, Ahmad M, MacLaughlin DT. Human Müllerian Inhibiting Substance: Enhanced purification imparts biochemical stability and restores antiproliferative effects. Protein Expression and Purification, 1992; 3(3):236-45.
Gustafson ML, Lee MM, Asmundson L, MacLaughlin DT, Donahoe PK. Mullerian Inhibiting Substance in the diagnosis and management of intersex and gonadal abnormalities. J Pedi Surg, 1993; 28:439-44.
Suen H-C, Catlin EA, Ryan DP, Wain JC, Donahoe PK. Biochemical immaturity of lungs in congenital diaphragmatic hernia. J Pedi Surg, 1993; 28:471-7.
He WW, Gustafson ML, Hirobe S, Donahoe PK. The developmental expression of four novel serine/threonine kinase receptors homologous to the Activin/TGF-b II receptor family. Developmental Dynamics, 1993; 196(2):133-42.
Bassing C, Yingling J, Wang T, Howe D, He WW, Gustafson M, Shah P, Donahoe P, Wang X-F. A Transforming Growth Factor Beta that Signals to Activate Gene Expression. Science, 1994; 263:87-9.
Suen HC, Bloch KD, Donahoe PK. Antenatal glucocorticoid treatment corrects pulmonary immaturity in experimentally induced congenital diaphragmatic hernia in rats. Pediatric Research 1994; 35:523-9.
Teixeira J, He WW, Shah PC, Morikawa N, Lee MM, Catlin EA, Hudson PL, Wing J, MacLaughlin DT, Donahoe PK. Developmental expression of a candidate Mullerian Inhibiting Substance type II receptor. Endocrinology, 1996; 137:160-65.
Lee, MM, Donahoe, P.K., Hasegawa, T., Silverman, B., Crist, G.B., Best, S., Hasegawa, Y., Noto, R.A., Schoenfeld, D., MacLaughlin, D.T. "Müllerian Inhibiting Substance (MIS) in humans: Normal levels from infancy to adulthood, The Journal of Clinical Endocrinology & Metabolism, 1996; 81:571-76.
Wang, TW, Li, BY, Danielson PD, Shah PC, Rockwell S, Lechleider RJ, Martin J, Manganaro T, Donahoe PK. The immunophilin FKBP12 functions as a common inhibitor of the TGF-b family type I receptors. Cell 1996; 86: 435-44.
Hedrick HL, Kaban JM, Pacheco BA, Losty PD, Doody DP, Ryan DP, Manganaro TF, Donahoe PK, Schnitzer JJ. Prenatal glucocorticoids improve pulmonary morphometrics in fetal sheep with congenital diaphragmatic hernia. J Pediatr Surg. 1997; 32(2):217-222.
Gu Z, Reynolds EM, Song J, Lei H, Donahoe PK, Li E. The type I serine-threonine kinase receptor ActRIA functions in extraembryonic cells to regulate mouse gastrulation. Development 1999 Jun;126(11):2551-61.
Islam S, Narra V, Cote GM, Manganaro TF, Donahoe PK, Schnitzer JJ. Prenatal vitamin E treatment improves lung growth in fetal rats with congenital diaphragmatic hernia. Journal of Pediatric Surgery 1999 Jan;34 (1):172-6.
Lane AH, Lee MM, Fuller Jr. AF, Kehas DJ, Donahoe PK, MacLaughlin DT. Diagnostic utility of Müllerian Inhibiting Substance determination in patients with primary and recurrent granulosa cell tumors. Gynecologic Oncology, 1999 Apr;73(1):51-5.
Masiakos PT, MacLaughlin DT, Teixeira J, Maheswaran S, Shah PC, Kehas DJ, Kenneally MK, Dombkowski DM, Ha TU, Preffer FI, Donahoe PK. Human Ovarian Cancer, Cell Lines and Primary Ascites Cells, Express the Human MIS Type II Receptor, Bind, and are Responsive to MIS. Clin Cancer Res 1999 Nov; 5(11):3488-99.
Teixeira JM, Fynn-Thompson E, Payne AT, Donahoe PK. Mullerian-Inhibiting Substance Regulates Androgren Synthesis at the Transcriptional Level. Endocrinology 1999; 140(10): 4732-4738.
Ha TU, Segev DL, Barbie D, Masiakos PT, Tran TT, Dombkowski D, Glander M, Clarke T, Lorenzo HK, Donahoe PK, Maheswaran S. Müllerian Inhibiting Substance inhibits cell growth through a Rb independent mechanism. J Biol Chem. 2000 Nov; 275(47):37101-9.
Clarke TR, Hoshiya Y, Yi SE, Liu X, Lyons KM, and Donahoe PK. Müllerian Inhibiting Substance Signaling uses a BMP-like pathway mediated by ALK2 and induces Smad6 expression. Mol Endocrinol. 2001 June; 15(6): 946-959.
Segev DL, Hoshiya Y, Hoshiya M, Tran TT, Carey JL, Stephen AE, MacLaughlin DT, Donahoe PK, Maheswaran S. Mullerian-Inhibiting Substance regulates NF-kB signaling in the prostate in vitro and in vivo. PNAS 2002;99(1):239-244.
Kling DE, Lorenzo HK, Trbovich AM, Kinane TB, Donahoe PK, Schnitzer JJ. MEK-1/2 inhibition reduces branching morphogenesis and causes mesenchymal cell apoptosis in fetal rat lungs. Am J Physiol. 2002; 282: L370-L378.
Stephen AE, Pearsall LA, Christian BP, Donahoe PK, Vacanti JP, MacLaughlin DT. Highly purified Mullerian Inhibiting Substance inhibits ovarian cancer in vivo. Clinical Cancer Research 2002; 8:2640-2646.
Laurich VM, Trbovich AM, O'Neill FH, Houk CP, Payne AH, Donahoe PK, Teixeira J. Mullerian Inhibiting Substance blocks the PKA-induced expression of cytochrome P450C17-20 hyrdroxylase/lase mRNA in mouse Leydig cells independent of CREB phosphorylation. Endocrinology 2002; 143:3351-3360.
Lorenzo HK, Teixeira J, Pahlavan N, Laurich VM, Donahoe PK, MacLaughlin DT. New approaches for high-yield purification of Müllerian inhibiting substance improve its bioactivity. J Chromatogr B 2002; 766:89-98.
Misra M, MacLaughlin DT, Donahoe PK, and Lee, MM. The Role of Mullerian-Inhibiting Substance in the Evaluation of Phenotypic Female Patients with Mild Degrees of Virilization. J Clin Endo & Met 2003; 88(2)787-92.
Misra M, MacLaughlin D, Donahoe P, and Lee MM. Measurement of MIS facilitates management of boys with miscrophalius and cryptorchidism. J Clin Endocrinol Metab 2002; 87:3598-3602.
Hoshiya M. Christian B., Cromie W., Zhan Y, MacLaughlin DT, and Donahoe PK. Persistent Mullerian Duct Syndrome Caused by Both a 27-bp Deletion and a Novel Splicing Mutation in the MIS type II Receptor Gene. Birth Defects Research (Part A) 2003; 67:868-874.
Dasgupta R, Schnitzer JJ, Hendren WH, and Donahoe PK. Congenital Adrenal Hyperplasia: Surgical Considerations of a 46XX Patient Raised as a Male. J Pedi Surg 2003;38(8):1269-1273.
Hoshiya Y, Gupta V., Kawakubo H, Brachtel E, Carey J, Sasur L, Scott A, Donahoe PK, and Maheswaran S. MIS Promotes INF-g-induced gene expression and apoptosis in breast cancer cells. JBC 2003; 278(51):51703-51712.
Barbie TU, Barbie DA, MacLaughlin DT, Maheswaran S, and Donahoe PK. Mullerian Inhibiting Substance inhibits cervical cancer cell growth via a pathway involving p130 and p107. PNAS 2003;100(26):15601-15606.
MacLaughlin DT, and Donahoe PK. Sex Determination and Differentiation. NEJM 2004; 350(4):367-378.
Houk CP, Pearson EJ, Martinelle N, Donahoe PK, Teixeira J. Feedback Inhibition of Steroidogenic Acute Regulatory Protein Expression in Vitro and In Vivo by Androgens. Endocrinology 2004; 145(3):1269-1275.
Salva A, Hardy MP, Wu XF, Sottas CM, MacLaughlin DT, Donahoe PK, Lee MM. Mullerian Ihibiting Substance Inhibits Rat Leydig Cell Regeneration after Ethylene Dimethanesulphonate Ablation. Bio of Repro 2004;70:600-607.
Houk CP and Donahoe PK. Mullerian Inhibiting Substance: New Insights. Encylopedia of Endo Diseases 2004;3;268-270.
Donahoe PD. Sustained inquiry: in the clinic and at the bench. J Pedi Surg 2004;39(11):1601-6.
Iafrate AJ, Feuk L, Rivera MN, Listewnik ML, Donahoe PK, Qi Ying, Scherer SW, Lee C. Detection of large-scale variation in the human genome. Nature Genetics 2004;1-3.
Arango NA, Pearson EJ, Donahoe PK, Teixeira J. Genomic structure and expression analysis of mouse testis-specific ribbon protein (Trib). Gene 2004;343:221-227.
Trbovich AM, Martinelle N, O’Neill FH, Pearson EJ, Donahoe PK, Sluss PM, Teixeira J. Steroidogenic activities in MA-10 Leydig cells are differentially altered by cAMP and Mullerian Inhibiting Substance. J Steroid Bio and Mol Bio 2004;92:199-208.
Stoler JM, Leach NT, Donahoe PK. Case 36-2004: A 23-day old infant with hypospadias and failure to thrive. NEJM 2004;351:2319-26.
Kling DE, Aidlen JT, Fisher JC, Kinane TB, Donahoe PK, and Schnitzer JJ. Nitrofen induces a redox-dependent apoptosis associated with increased p38 activity in P19 teratocarcinoma cells. Toxicology in Vitro 2005;19(1):1-10.
Renaud EJ, Oliva E, Rueda B, MacLaughlin DT, Donahoe, PK. Endometrial cancer is a receptor mediated target for Mullerian Inhibiting Substance. PNAS 2005;102(1):111-6.
Gupta V, Carey J, Kawakubo H, Muzikansky A, Green J, Donahoe PK, MacLaughlin DT, Maheswaran S. Mullerian Inhibiting Substance suppresses tumor grown in the C3(1)T antigen transgenic mouse mammary carcinoma model. PNAS 2005; 102(9):3219-24.
Klaassens M, van Dooren M, Eussen HJ, Douben H,DekkerX,Lee C, Donahoe PK, Galijaard RJ, Goemaere N,deKrijger RR, Wouters C,Wauters J, Oostra BA, Tibboel D, deKlein A. Congenital Diaphragmatic Hernia and Chromosome 15q26: determination of a candidate region by use of fluorescent in situ hydridization and array-based comparative genomic hybirization. Amer J of Human Genetics 2005; 76:000-000.
Kawaguchi AL, Donahoe PK, Ryan DP. Management and long-term follow-up of patients with type III and IV laryngotracheoesophageal clefts. J Pedi Surg 2005; 40:158-165.
Arango NA, Szotek PP, Manganaro TF, Ester Oliva Donahoe PK, Teixeira JT. Conditional deletion of b-catenin in the mesenchyme of the developing mouse uterus results in a cell fate determinatin switch from myogenesis to adipogenesis. Dev Biol in press 2005.
Kantarci S, Casavant D, Prada C, Russell M, Byrne J, Wilkins Haug L, Jennings R, Manning S, Blaise JF, Boye TK, Fryns JP, Holmes LB, Donahoe PK, Lee, C, Kimonis V, Pober B. Findings from aCGH in patients with congenital diaphragmatic hernia (CDH): a possible locus for Fryns syndrome. Am J Med Genetics 2005 (submitted).
Pieretti-Vanmarcke R, Donahoe PK, Szotek P, Manganaro T, Lorenzen MK, Lorenzen J, Connolly DC, Halpern EF, MacLaughlin DT. Recombinant Human MIS inhibits MISRII-directed Transgenic Mouse Ovarian Cancers in a Preclinical in vivo Model. Submitted 2005.
Zhan Y, MacLaughlin DT, Manganaro TF, Szotek PP, Arango NA, Teixeira J, Donahoe PK. Mullerian Inhibiting Substance regulates its receptor/Smad signaling and causes mesenchymal transition of the coelomic epithelial cells early in Mullerian duct regression. Submitted 2005.
Wang PY, Koish K, McGeachie AB, Kimber M, MacLaughlin DT, Donahoe PK, McLennan I. Mullerian Inhibiting Substance acts as a novel motor neuron survival factor in vivo. PNAS. In press.
Kling DE, Brandon KL, Sollinger CA, Cavicchio AJ, Qingyuan GE, Kinane TB, Donahoe PK, Schnitzer JS. Distribution of ERK1/2 and ERK3 during normal rat fetal lung development. Anat and Embryo. In Press 2005.
