Current Environment:

 

Ongoing Projects

Identification of Therapeutics Targeting Endometriosis

Endometriosis is a chronic illness characterized by infertility and/or chronic pain that results from ectopic growth of endometrial tissue in affected women.  Endometriotic lesions are present in >50% of women with chronic pelvic pain, and in a similar fraction of infertile women.  Definitive diagnosis requires surgical visualization, and as a result is often delayed—on average, seven years elapse between the onset of symptoms and diagnosis.  Available therapies include pain management, hormonal manipulation and surgery.  However, the disorder is typically progressive, and none of these options is entirely effective in the long-term.  New approaches to endometriosis treatment are urgently needed.

To identify new therapeutics for endometriosis, we are currently working to Establish and validate a mouse model of endometriosis-associated pain.  We use allotransplanted endometrial tissue to induce lesions and then use well-validated outcome measures to determine local mechanical hypersensitivity (von Frey stimulation) as well as a non-reflexive measures of pain (thermal gradient and behavioral changes.)  Ongoing work to validate this model involves treatment with therapeutics known to alleviate pain in endometriosis sufferers, with future work focused on identifying FDA-approved drugs with known or hypothesized activity in treating pain or angiogenesis that can be repurposed to treat endometriosis.  We focus on compounds that are well-tolerated and safe in pregnancy (category A or B).  Those found to be effective in the  animal model can then be rapidly transitioned into clinical trials in collaboration with the Boston Center for Endometriosis (BCE).

Identifying Antagonists of Antizyme Inhibitor  

Currently, very few cancer therapies result in long-term tumor stabilization or regression.  This may be because most current therapies target a relatively small number of proteins and pathways.  For this reason, we have developed evidence validating antizyme inhibitor (AZI) as a novel therapeutic target.  AZI gene amplification and overexpression are common in a variety of tumor types, including cancers of the prostate, breast, ovary, testis, liver, lung, and skin.  In addition, we have found that AZI silencing suppresses cell proliferation and experimental cancer in animal models.  AZI promotes tumor growth by several pathways.  These include binding and sequestering the ornithine decarboxylase (ODC) antagonist antizyme (AZ), inhibiting polyamine uptake, and inactivating ODC, the rate-limiting enzyme in polyamine synthesis.  AZ sequestration also derepresses cyclin D1, SMAD1, and aurora A kinase.  Inhibition of only one of these (ODC) with DFMO has resulted in anticancer activity in clinical trials, but its success has been modest, likely because of a compensatory increase in uptake of extracellular polyamines that is blocked by AZ.  Because of the increased range of oncogenic activities regulated by AZ, molecules that increase available intracellular AZ by blocking AZI are expected to be more active than DFMO.  Such molecules have not yet been discovered; however, they are likely to exist because AZI retains a pocket analogous to the known substrate-binding pocket in ODC to which it is homologous.  Given the overlap between this pocket and the AZ binding site, small molecule inhibitors represent an opportunity to block AZ binding and sequestration and increase available AZ in the cell. 

In collaboration with the Zetter lab, we are working to develop a drug discovery pipeline designed to identify molecules that inhibit the AZ-AZI interaction.  The first stage of this pipeline will be high throughput FRET assays to identify inhibitors of AZ-AZI interaction, using AZ-ODC and AZ-AZIN2 FRET interaction assays as counterscreens.  Secondary assays to assess the ability of hit compounds to disrupt AZ-AZI complexes in vivo will include BLI (BioLayer Interferometry) and in vivo FRET assays to assess AZ-AZI interaction in cultured cells.  This pipeline will be tested at the Institute of Chemistry and Cell Biology (ICCB) facility at Harvard Medical School in Boston.  Small molecule AZI antagonists identified in the proposed screening pipeline will be used as lead compounds for the design of anticancer therapeutics and as probes of the molecular recognition and cellular effects of AZ release.  

  • Ghalali A, Rice JM, Kusztos A, Jernigan FE, Zetter BR, Rogers MS.  Developing a novel FRET assay, targeting the binding between Antizyme-AZIN.  Sci Rep. 2019;15;9:4632.

The Role of the Anthrax Toxin Receptors in Angiogenesis.

I collaborated with Dr. Kenneth Christensen and Dr. John Collier to demonstrate that the Protective Antigen subunit of anthrax toxin is a potent anti-angiogenic agent (a).  This establishes the anthrax toxin receptors as new targets for antiangiogenic therapy.  Dr. Christensen and I are continuing this collaboration to clarify the role of each of the two anthrax toxin receptors in angiogenesis.  This has included collaborations with the National Screening Laboratory for the Regional Centers of Excellence for Biodefense and Emerging Infectious Diseases (NSRB) facility to develop assays for small molecule inhibitors of these molecules (b, c).  Screening of natural product libraries in collaboration with Dr. Jon Clardy has led to the identification of two different classes of inhibitors for CMG2 (d and Cao, et al.) and ongoing work also has identified hits from synthetic small molecule libraries for both CMG2 and TEM8.  Such molecules are likely to serve as good leads for both anthrax-protective agents as well as antiangiogenic agents. 

  • Rogers MS, Christensen KA, Birsner AE, Short SM, Wigelsworth DJ, Collier RJ, D’Amato RJ.Mutant anthrax toxin B-moiety (protective antigen) inhibits angiogenesis and tumor growth.Cancer Research 2007;67:9980-5.
  • Rogers MS, Cryan LM, Habeshian KA, Bazinet L, Caldwell TP, Ackroyd PC, Christensen KA.A FRET-based high throughput screening assay to identify inhibitors of anthrax protective antigen binding to capillary morphogenesis gene 2 protein.PLoS One 2012;7:e39911. PMCID: PMC3386954.
  • Cryan L, Habeshian K, Caldwell TP, Morris MT, Ackroyd PC, Christensen KA, Rogers MS.Identification of small molecules that inhibit the interaction of TEM8 with anthrax protective antigen using a FRET assay.Journal of Biomolecular Screening 2013;18:714-25. PMCID: PMC3859190.
  • Cryan LM, Bazinet L, Habeshian KA, Cao S, Clardy J, Christensen KA, Rogers MS.1,2,3,4,6-penta-O-galloyl-β-D-glucopyranose inhibits angiogenesis via inhibition of capillary morphogenesis gene 2.Journal of Medicinal Chemistry 2013;56:1940-5. PMCID: PMC3600088.

Genetic Differences that Affect the Host Response to Angiogenic Stimuli.

At the outset of these studies, dramatic (>10-fold) differences in the corneal neovascular response to bFGF had been observed among different inbred mouse strains, but the genetic structure of this trait and its relationship to possible differences in angiogenesis and other endothelial phenotypes in other tissues was unclear.  We used recombinant inbred mouse strains to determine that corneal angiogenic response to bFGF is a complex (i.e. multigenic) trait and identified several quantitative trait loci (QTLs) that control the response to bFGF in the cornea (a).  We found that the same is true of corneal response to VEGF (b) and laser-induced choroidal neovascularization—the latter being a model of age-related macular degeneration (c).  We also determined that there is substantial correlation among VEGF-induced corneal neovascularization, bFGF-induced corneal neovascularization, and laser-induced choroidal neovascularization, with overlapping loci identified in some cases.  This indicates that a substantial fraction of the variation in angiogenic responsiveness is shared among multiple ocular tissues and growth factors.  We further supported this idea by demonstrating that quantitative vessel outgrowth in the aortic ring assay, vessel growth in the matrigel plug assay, the number of circulating endothelioid cells (CECs), and the number of circulating endothelioid precursors (CEP) all correlate with corneal angiogenic response to bFGF and VEGF (d).  We have thus demonstrated genetic control of a core final common angiogenesis pathway and further identified polymorphic loci that regulate the angiogenic response to multiple physiologically important stimuli.  Further work to understand the role of these genes/pathways in cancer development is underway.

  • Rogers MS, Rohan RM, Birsner AE, D’Amato RJ.Genetic loci that control vascular endothelial growth factor-induced angiogenesis.FASEB Journal 2003;17:2112-4.
  • Rogers MS, Rohan RM, D’Amato RJ.Genetic loci that control the angiogenic response to basic fibroblast growth factor.FASEB Journal 2004;18:1050-9.
  • Nakai K*, Rogers MS*, Baba T, Funakoshi T, Birsner AE, Luyindula DS, D’Amato RJ.Genetic loci that control the size of laser-induced choroidal neovascularization.FASEB Journal 2009;23:2235-43. PMCID: PMC2704592. *These authors contributed equally to this work.
  • Shaked Y, Bertolini F, Man S, Rogers MS, Cervi D, Foutz T, Rawn K, Voskas D, Dumont DJ, Ben-David Y, Lawler J, Henkin J, Huber J, Hicklin DJ, D'Amato RJ, Kerbel RS.Genetic heterogeneity of the vasculogenic phenotype parallels angiogenesis; implications for cellular surrogate marker analysis of antiangiogenesis.Cancer Cell 2005;7:101-11.

Completed Projects

Antimyeloma Activity of the Thalidomide Analogs, Pomalidomide and Lenalidomide.

My early work at Boston Children’s Hospital focused on the identification of thalidomide analogs that exhibit improved anti-cancer activity.  At the initiation of my work, thalidomide had been shown to exhibit antiangiogenic and anti-TNFα activity and remarkable anti-tumor activity had recently been observed in multiple myeloma.  However, the mechanism by which the thalidomide affected myeloma was unclear.  In collaboration with a small company (EntreMed), we used structure-activity studies to identify amino and hydroxyl derivatives of thalidomide with increased direct anti-myeloma-cell activity (a); hydroxyl metabolites of thalidomide being likely responsible for the strong clinical activity observed against myeloma.  We further showed that the direct anti-myeloma cell activity is independent of previously identified biological activities in analog series.  Thus, we determined that a novel mechanism is responsible for the direct myeloma cell killing observed with amino analogs of thalidomide (b).  We then performed the first in vivo evaluation of these analogs in mouse models of multiple myeloma and showed that the combined antiangiogenic and anti-myeloma activity of S-3-aminothalidomide (enantiomerically pure pomalidomide) can cure experimental animals with established tumors.  This work was critical to the initiation of clinical trials, and the racemic mixture is now FDA-approved and is still the most active of the thalidomide analogs against multiple myeloma (c, d).

  • D’Amato RJ, Lentzsch S, Anderson KC, Rogers MS.Mechanism of action of thalidomide and 3-aminothalidomide in multiple myeloma.Seminars in Oncology 2001;28:597-601.
  • Lentzsch S*, Rogers MS*, LeBlanc R, Birsner AE, Shah JH, Treston AM, Anderson KC, D'Amato RJ.S-3-amino-phthalimido-glutarimide inhibits angiogenesis and growth of B-cell neoplasias in mice.Cancer Research 2002;62:2300-5. *These authors contributed equally to this work.
  • Folkman J and Rogers MS.Thalidomide for multiple myeloma.New England Journal of Medicine 2006;354:2389-90.
  • D’Amato RJ, Lentzsch S, Rogers MS.Pomalidomide is strongly anti-angiogenic and teratogenic in relevant animal models.Proceedings of the National Academy of Sciences, USA 2013;110(50):E4818. PMCID: PMC3864340.

The Role of Coat-Color Genes in the Regulation of Angiogenesis.

As outlined above, the magnitude of the angiogenic response to a given stimulus varies widely (>10-fold) among different inbred mouse strains.  However, the identities of genes affecting this trait were not known.  We used multiple F2 inbred strain crosses to identify genomic regions containing polymorphisms that affect the angiogenic response to bFGF in the corneal micropocket assay (a).  To identify VEGF response regulating polymorphisms we used the B6.A consomic strain series.  We followed up by generating congenic and subcongenic animals to narrow the region of interest.  Data from these animals, along with novel bioinformatics techniques, identified the albino allele as the polymorphism responsible for AngVq4 (b), and the pink-eyed dilution allele as responsible for AngFq5(c).  We found that the small molecule pigment precursors 3,4-dihydroxyphenlyalanine and 5,6-dihydroxyindole mediate a portion of the effects of these alleles.  We further discovered that a TGFβ regulator, fibromodulin, is differentially expressed in albino vs. pigmented melanocytes, exhibits differing circulating levels in albino and pigmented C57BL/6J animals and humans with differing skin pigmentation, and affects endothelial cell function in vitro and angiogenesis in vivo (d).  These studies identified novel, unexpected angiogenesis regulatory pathways and proteins, thereby demonstrating the power of genetic approaches to biological questions. 

  • Rogers MS, Birsner AE, D’Amato RJ.The mouse cornea micropocket angiogenesis assay.Nature Protocols 2007;2:2545-50.
  • Rogers MS†, Adini I, McBride AF, Birsner AE, D’Amato RJ.The albino mutation of tyrosinase alters ocular angiogenic responsiveness.Angiogenesis 2013;16:639-46. †Sole corresponding author
  • Rogers MS†, Boyartchuk V, Rohan RM, Birsner AE, Dietrich WF, D’Amato RJ.The classical pink-eyed dilution mutation affects angiogenic responsiveness.PLoS ONE 2012;7:e35237. PMCID: PMC3352893. †Sole corresponding author
  • Adini I, Ghosh K, Adini A, Zailong C, Yoshimura T, Benny O, Connor KM, Rogers MS, Bazinet L, Birsner AE, Bielenberg DR, D'Amato RJ.Melanocyte-secreted fibromodulin promotes an angiogenic microenvironment.Journal of Clinical Investigation 2014;124(1):425-36. PMCID: PMC3871226.