Salivary glands as a potential gene transfer target for gene therapeutics of some monogenetic endocrine disorders.

Salivary glands (SGs) exhibit several important features which are also common to endocrine glands: self-containment due to a surrounding capsule, highly efficient protein production and the ability to secrete proteins into the bloodstream. We have hypothesized that SGs are potentially useful as gene transfer targets for the correction of inherited monogenetic endocrine disorders. In the present communication, we extend our studies and attempt to test our hypothesis by comparing the efficacy of two commonly used viral vectors and the resulting serum and salivary distribution of transgene encoded hormones. A low dose (5 x10(9) particles) of either an adenoviral serotype 5 (Ad5) vector encoding the human erythropoietin (hEPO) cDNA or an adeno-associated virus sero-type 2 (AAV2) vector encoding either the hEPO or human growth hormone (hGH) cDNA was administered to individual submandibular SGs of Balb/c mice. Serum and salivary hEPO and hGH levels were determined at defined time points. Two additional recombinant viruses encoding enhanced green fluorescence protein (GFP) were also used (AdGFP and AAVGFP) in order to perform immunohistochemical analyses of transgenic protein localization in SG sections post-administration. AAV2 vectors led to stable gene transfer unlike the results with the Ad5 vectors. Indeed, in one mouse we observed hEPO production for a period of two years after administration of AAVhEPO to SGs. hEPO, which is a constitutive pathway secretory protein, was readily secreted into the bloodstream from the SGs, yielding therapeutically adequate serum levels. Conversely, hGH, a regulated secretory pathway protein, was preferentially secreted into saliva. SGs may be an attractive candidate target tissue for gene therapeutics of some monogenetic endocrine deficiency disorders. At present, AAV2 vectors seem particularly useful for such applications, and transgenes encoding constitutive secretory pathway hormones are more suitable for this application with SGs than those encoding regulated secretory pathway hormones.

[1]  美昭 森山,et al.  Erythropoietin , 1972, Reactions Weekly.

[2]  S. Yu,et al.  Repetitive adenovirus administration to the parotid gland: role of immunological barriers and induction of oral tolerance. , 1998, Human gene therapy.

[3]  J W Fisher,et al.  Glycosylation at specific sites of erythropoietin is essential for biosynthesis, secretion, and biological function. , 1988, The Journal of biological chemistry.

[4]  A. Castle,et al.  Intracellular transport and secretion of salivary proteins. , 1998, Critical reviews in oral biology and medicine : an official publication of the American Association of Oral Biologists.

[5]  S. Mujais,et al.  Erythropoietin is produced by tubular cells of the rat kidney , 2007, Cell Biochemistry and Biophysics.

[6]  Ronald G. Crystal,et al.  The gene as the drug , 1995, Nature Medicine.

[7]  P. Dannies.  Mechanisms for storage of prolactin and growth hormone in secretory granules. , 2002, Molecular genetics and metabolism.

[8]  D. Steiner,et al.  A mutation in the B chain coding region is associated with impaired proinsulin conversion in a family with hyperproinsulinemia. , 1987, Proceedings of the National Academy of Sciences of the United States of America.

[9]  D. Bohl,et al.  Mechanisms leading to sustained reversion of beta-thalassemia in mice by doxycycline-controlled Epo delivery from muscles. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[10]  S. Rothman,et al.  The endocrine secretion of mammalian digestive enzymes by exocrine glands. , 1999, The American journal of physiology.

[11]  R. Kotin,et al.  High-efficiency transfer of the T cell co-stimulatory molecule B7-2 to lymphoid cells using high-titer recombinant adeno-associated virus vectors. , 1995, Human gene therapy.

[12]  J. M. González,et al.  Engineering the largest RNA virus genome as an infectious bacterial artificial chromosome. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[13]  G. Veres,et al.  Intravenous administration of an AAV-2 vector for the expression of factor IX in mice and a dog model of hemophilia B , 2004, Gene Therapy.

[14]  I. Bossis,et al.  Reengineered salivary glands are stable endogenous bioreactors for systemic gene therapeutics. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[15]  L. Johnson,et al.  Physiology of the gastrointestinal tract , 2012 .

[16]  T. Clackson,et al.  Regulated expression of erythropoietin from an AAV vector safely improves the anemia of beta-thalassemia in a mouse model. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[17]  J. Bazan.  A novel family of growth factor receptors: a common binding domain in the growth hormone, prolactin, erythropoietin and IL-6 receptors, and the p75 IL-2 receptor beta-chain. , 1989, Biochemical and biophysical research communications.

[18]  P. Aebischer,et al.  Salivary glands and gene therapy: the mouth waters , 2004, Gene Therapy.

[19]  B. Baum,et al.  Rapamycin control of exocrine protein levels in saliva after adenoviral vector-mediated gene transfer , 2004, Gene Therapy.

[20]  B. Baum,et al.  Immediate inflammatory responses to adenovirus-mediated gene transfer in rat salivary glands. , 1996, Human gene therapy.

[21]  G. Palù,et al.  New perspectives for gene therapy in endocrinology. , 2000, European journal of endocrinology.

[22]  B. Trapnell,et al.  Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy , 1996, Journal of virology.

[23]  B. Baum,et al.  Gene transfer to salivary glands. , 2002, International review of cytology.

[24]  Y. Marmary,et al.  Systemic action of human growth hormone following adenovirus-mediated gene transfer to rat submandibular glands , 1998, Gene Therapy.

[25]  B. Baum,et al.  Evaluation of salivary gland acinar and ductal cell-specific promoters in vivo with recombinant adenoviral vectors. , 2001, Human gene therapy.

[26]  A V Nieuw Amerongen,et al.  Saliva--the defender of the oral cavity. , 2002, Oral diseases.

[27]  P. Rakoczy,et al.  Adenovirus and adeno-associated virus vectors. , 2002, DNA and cell biology.

[28]  R. Kelly.  Pathways of protein secretion in eukaryotes. , 1985, Science.

[29]  B. Baum,et al.  Polarized secretion of transgene products from salivary glands in vivo. , 1999, Human gene therapy.

[30]  Lili Wang,et al.  Noninvasive gene transfer to the lung for systemic delivery of therapeutic proteins. , 2002, The Journal of clinical investigation.

[31]  J. Chiorini,et al.  Scalable purification of adeno-associated virus type 2, 4, or 5 using ion-exchange chromatography. , 2002, Human gene therapy.

[32]  A. Hoque,et al.  Hydroxychloroquine enhances the endocrine secretion of adenovirus-directed growth hormone from rat submandibular glands in vivo. , 2001, Human gene therapy.

[33]  H. Gerdes,et al.  Signal-mediated sorting to the regulated pathway of protein secretion. , 1999, Annals of anatomy = Anatomischer Anzeiger : official organ of the Anatomische Gesellschaft.

[34]  R. Samulski,et al.  Sustained and complete phenotype correction of hemophilia B mice following intramuscular injection of AAV1 serotype vectors. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[35]  Jianghuai Wang,et al.  Utilizing endocrine secretory pathways in salivary glands for systemic gene therapeutics , 2004, Journal of cellular physiology.

[36]  K. Csaky,et al.  Saturation of, and competition for entry into, the apical secretory pathway. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[37]  R. Hammer,et al.  A mutant human proinsulin is secreted from islets of Langerhans in increased amounts via an unregulated pathway. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[38]  Luigi Naldini,et al.  Viral vectors for gene therapy: the art of turning infectious agents into vehicles of therapeutics , 2001, Nature Medicine.

[39]  G. Goldspink.  Skeletal muscle as an artificial endocrine tissue. , 2003, Best practice & research. Clinical endocrinology & metabolism.

[40]  N. Cawley,et al.  Mechanism of Sorting Proopiomelanocortin and Proenkephalin to the Regulated Secretory Pathway of Neuroendocrine Cells , 2002, Annals of the New York Academy of Sciences.

[41]  D. Bohl,et al.  Improvement of erythropoiesis in beta-thalassemic mice by continuous erythropoietin delivery from muscle. , 2000, Blood.

[42]  A. Gown,et al.  Persistent and therapeutic concentrations of human factor IX in mice after hepatic gene transfer of recombinant AAV vectors , 1997, Nature Genetics.

[43]  R. Kotin,et al.  Recombinant adeno-associated virus serotype 2 vectors mediate stable interleukin 10 secretion from salivary glands into the bloodstream. , 2002, Human gene therapy.

[44]  S. Pillemer,et al.  Local adeno-associated virus-mediated interleukin 10 gene transfer has disease-modifying effects in a murine model of Sjögren's syndrome. , 2003, Human gene therapy.