Ann Richmond

Vanderbilt University, Нашвилл, Michigan, United States

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Publications (121)722.04 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Preclinical studies show that Inhibition of aurora kinases in melanoma tumors induces senescence and reduces tumor growth, but does not cause tumor regression. Additional preclinical models are needed to identify agents that will synergize with aurora kinase inhibitors to induce tumor regression. We combined treatment with an aurora kinase A inhibitor, MLN8237, with agents that activate death receptors (Apo2L/TRAIL or death receptor 5 agonists) and monitored the ability of this treatment to induce tumor apoptosis and melanoma tumor regression using human cell lines and patient derived xenograft mouse models. We found that this combined treatment led to apoptosis and markedly reduced cell viability. Mechanistic analysis showed that the induction of tumor cell senescence in response to the AURKA inhibitor resulted in a decreased display of Apo2L/TRAIL decoy receptors and increased display of one Apo2L/TRAIL receptor (death receptor 5), resulting in enhanced response to death receptor ligand/agonists. When death receptors were activated in senescent tumor cells, both intrinsic and extrinsic apoptotic pathways were induced independent of BRAF, NRAS or p53 mutation status. Senescent tumor cells exhibited BID- mediated mitochondrial depolarization in response to Apo2L/TRAIL treatment. In addition, senescent tumor cells had a lower apoptotic threshold due to decreased XIAP and survivin expression. Melanoma tumor xenografts of human cell lines or patient derived xenografts displayed treatmentsignificantly improved treatment. These findings provide a strong rationale for combining senescence-inducing therapeutics with death receptor agonists for improved cancer treatment. Copyright © 2015, American Association for Cancer Research.
    Clinical Cancer Research 07/2015; DOI:10.1158/1078-0432.CCR-15-0293 · 8.19 Impact Factor
  • Anna Vilgelm, Ann Richmond
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    ABSTRACT: We recently demonstrated that therapy combining Aurora Kinase A and MDM2 antagonists is effective against melanoma in preclinical settings. Notably, besides inducing apoptosis, this regimen led to tumor senescence and stimulated the host's anti-tumor immune defenses. Treatments leveraging both cancer cell-intrinsic and extrinsic anti-tumor mechanisms can improve melanoma therapeutic outcomes.
    OncoImmunology 05/2015; DOI:10.1080/2162402X.2015.1009299 · 6.28 Impact Factor
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    ABSTRACT: Nuclear LASP-1 (LIM and SH3 protein-1) has a direct correlation with overall survival of breast cancer patients. In this study, immunohistochemical analysis of a human breast TMA showed that LASP-1 is absent in normal human breast epithelium but the expression increases with malignancy and is highly nuclear in aggressive breast cancer. We investigated whether the chemokines and growth factors present in the tumor microenvironment could trigger nuclear translocation of LASP-1.Treatment of human breast cancer cells with CXCL12, EGF and HRG, and HMEC-CXCR2 cells with CXCL8 facilitated nuclear shuttling of LASP-1. Data from the biochemical analysis of the nuclear and cytosolic fractions further confirmed the nuclear translocation of LASP-1 upon chemokine and growth factor treatment. CXCL12-dependent nuclear import of LASP-1 could be blocked by CXCR4 antagonist, AMD-3100. Knock down of LASP-1 resulted in alterations in gene expression leading to an increased level of cell-junction and extracellular matrix proteins and an altered cytokine secretory profile. Three-dimensional cultures of human breast cancer cells on Matrigel revealed an altered colony growth, morphology and arborization pattern in LASP-1 knockdown cells. Functional analysis of the LASP-1 knockdown cells revealed increased adhesion to collagen IV and decreased invasion through the Matrigel. Proteomic analysis of immunoprecipitates of LASP-1 and subsequent validation approaches revealed that LASP-1 associated with the epigenetic machinery especially UHRF1, DNMT1, G9a and the transcription factor Snail1. Interestingly, LASP-1 associated with UHRF1, G9a, Snail1 and di- and tri-methylated histoneH3 in a CXCL12-dependent manner based on immunoprecipitation and proximity ligation assays. LASP-1 also directly bound to Snail1 which may stabilize Snail1. Thus, nuclear LASP-1 appears to functionally serve as a hub for the epigenetic machinery.Oncogene advance online publication, 18 May 2015; doi:10.1038/onc.2015.166.
    Oncogene 05/2015; DOI:10.1038/onc.2015.166 · 8.56 Impact Factor
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    ABSTRACT: Therapeutics that induce cancer cell senescence can block cell proliferation and promote immune rejection. However, the risk of tumor relapse due to senescence escape may remain high due to the long lifespan of senescent cells that are not cleared. Here we show how combining a senescence-inducing inhibitor of the mitotic kinase Aurora A (AURKA) with an MDM2 antagonist activates p53 in senescent tumors harboring wildtype 53. In the model studied, this effect is accompanied proliferation arrest, mitochondrial depolarization, apoptosis and immune clearance of cancer cells by antitumor leukocytes in a manner reliant upon CCL5, CCL1 and CXCL9. The AURKA/MDM2 combination therapy shows adequate bioavailability and low toxicity to the host. Moreover, the prominent response of patient-derived melanoma tumors to co-administered MDM2 and AURKA inhibitors offers a sound rationale for clinical evaluation. Taken together, our work provides a preclinical proof-of-concept for a combination treatment which leverages both senescence and immune surveillance to therapeutic ends.
    Cancer Research 11/2014; 75(1). DOI:10.1158/0008-5472.CAN-14-2405 · 9.28 Impact Factor
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    ABSTRACT: Myeloid cells are effectors of both anti-tumor and pro-tumor immune responses, but much needs to be determined as to signals that determine which function of the myeloid lineage dominates. Shown here, mice with myeloid-specific IKKβ loss exhibit more rapid growth of cutaneous and lung melanoma tumors. Specifically, in a BRAF(V600E)PTEN-/- allograft model, IKKβ loss in macrophages resulted in reduced recruitment of myeloid cells into the tumor, reduced expression of MHCII, and enhanced production of the chemokine, CCL11, which negatively regulated dendritic cell maturation. The elevated serum and tissue levels of CCL11 mediated suppression of dendritic cell differentiation/maturation within the TME, resulted in a Th2 skew of the immune response, and impaired CD8+T cell-mediated tumor cell killing. Macrophage depletion or CD8+T cell depletion in mice with IKKβWT myeloid cells enhanced tumor growth in the melanoma allograft. In contrast, mice with IKKβWT myeloid cells used the myeloid cell response to mediate anti-tumor immunity against the syngeneic B16 melanoma, with less apparent dependency on a CD8+T cell response. Myeloid cells deficient in IKKβ were compromised in tumor cell killing based upon reduced ability to phagocytize and digest tumor cells. Conversely, mice with continuous IKKβ signaling in myeloid-lineage cells (IKKβCA) exhibited enhanced anti-tumor immunity and reduced B16 melanoma tumor growth. Collectively, these data uncover new mechanisms by which NF-κB signaling in myeloid cells promotes innate tumor surveillance.
    Cancer Research 10/2014; 74(24). DOI:10.1158/0008-5472.CAN-14-1091 · 9.28 Impact Factor
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    ABSTRACT: Stromal cells in the tumor microenvironment play a key role in the metastatic properties of a tumor. It is recognized that cancer-associated fibroblasts (CAFs) and endothelial cells secrete factors capable of influencing tumor cell migration into the blood or lymphatic vessels. We developed a microfluidic device that can be used to image the interactions between stromal cells and tumor cell spheroids in a three dimensional (3D) microenvironment while enabling external control of interstitial flow at an interface, which supports endothelial cells. The apparatus couples a 200-μm channel with a semicircular well to mimic the interface of a blood vessel with the stroma, and the design allows for visualization of the interactions of interstitial flow, endothelial cells, leukocytes, and fibroblasts with the tumor cells. We observed that normal tissue-associated fibroblasts (NAFs) contribute to the "single file" pattern of migration of tumor cells from the spheroid in the 3D microenvironment. In contrast, CAFs induce a rapid dispersion of tumor cells out of the spheroid with migration into the 3D matrix. Moreover, treatment of tumor spheroid cultures with the chemokine CXCL12 mimics the effect of the CAFs, resulting in similar patterns of dispersal of the tumor cells from the spheroid. Conversely, addition of CXCL12 to co-cultures of NAFs with tumor spheroids did not mimic the effects observed with CAF co-cultures, suggesting that NAFs produce factors that stabilize the tumor spheroids to reduce their migration in response to CXCL12.
    Biomicrofluidics 07/2014; 8(4):044105. DOI:10.1063/1.4890330 · 3.77 Impact Factor
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    ABSTRACT: Purpose To evaluate the efficacy and tolerability of bortezomib in combination with doxorubicin in patients with advanced hepatocellular carcinoma, and to correlate pharmacodynamic markers of proteasome inhibition with response and survival. Experimental Design This phase II, open-label, multicenter study examined the efficacy of bortezomib (1.3 mg/m(2) IV on d1, 4, 8, 11) and doxorubicin (15 mg/m(2) IV on d1, 8) in 21-day cycles. The primary endpoint was objective response rate. Results Best responses in 38 treated patients were 1 partial response (2.6 %), 10 (26.3 %) stable disease, and 17 (44.7 %) progressive disease; 10 patients were unevaluable. Median PFS was 2.2 months. Median OS was 6.1 months. The most common grade 3 to 4 toxicities were hypertension, glucose intolerance, ascites, ALT elevation, hyperglycemia and thrombosis/embolism. Worse PFS was seen in patients with elevated IL-6, IL-8, MIP-1α and EMSA for NF-κB at the start of treatment. Worse OS was seen in patients with elevated IL-8 and VEGF at the start of treatment. Patients had improved OS if a change in the natural log of serum MIP-1α/CCL3 was seen after treatment. RANTES/CCL5 levels decreased significantly with treatment. Conclusions The combination of doxorubicin and bortezomib was well-tolerated in patients with hepatocellular carcinoma, but the primary endpoint was not met. Exploratory analyses of markers of proteasome inhibition suggest a possible prognostic and predictive role and should be explored further in tumor types for which bortezomib is efficacious.
    Investigational New Drugs 06/2014; 32(5). DOI:10.1007/s10637-014-0111-8 · 2.93 Impact Factor
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    ABSTRACT: The chemokine receptor CXCR2 is vital for inflammation, wound healing, angiogenesis, cancer progression, and metastasis. Adaptor protein 2 (AP2), a clathrin binding heterotetrameric protein comprised of α, β2, μ2, and σ2 subunits, facilitates clathrin-mediated endocytosis. Mutation of the LLKIL motif in the CXCR2 carboxyl-terminal domain (CTD) results in loss of AP2 binding to the receptor and loss of ligand mediated receptor internalization and chemotaxis. AP2 knockdown also results in diminished ligand-mediated CXCR2 internalization, polarization and chemotaxis. Using knockdown/rescue approaches with AP2-μ2 mutants, the binding domains were characterized in reference to CXCR2 internalization and chemotaxis. When in an open conformation, μ2 Patch 1 and Patch 2 domains bind tightly to membrane PIP2 phospholipids. When AP2-μ2 is replaced with μ2 mutated in Patch 1 and/or Patch 2 domains, ligand-mediated receptor binding and internalization are not lost. However chemotaxis requires AP2-μ2 Patch 1, but not Patch 2. AP2-σ2 has been demonstrated to bind dileucine motifs to facilitate internalization. Expression of AP2-σ2 V88D and V98S dominant negative mutants resulted in loss of CXCR2 mediated chemotaxis. Thus, AP2 binding to both membrane phosphatidylinositol phospholipids and dileucine motifs is crucial for directional migration or chemotaxis. Moreover, AP2-mediated receptor internalization can be dissociated from AP2-mediated chemotaxis.
    Traffic 01/2014; 15(4). DOI:10.1111/tra.12154 · 4.71 Impact Factor
  • Yingjun Su, Ann Richmond
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    ABSTRACT: Significance: Efficient recruitment of neutrophils to an injured skin lesion is an important innate immune response for wound repair. Defects in neutrophil recruitment lead to impaired wound healing. Recent Advances: Chemokines and chemokine receptors are known to regulate neutrophil recruitment. Recent research advances reveal more mechanistic details about the regulation of chemokines and chemokine receptors on neutrophil egress from bone marrow, transmigration into the wound site, spatial navigation toward the necrotic skin tissue, and apoptosis-induced clearance by efferocytosis. Critical Issues: Skin injury triggers local and systemic alterations in the expression of multiple chemotactic molecules and the magnitude of chemokine receptor-mediated signaling. The responses of a number of CXC and CX3C chemokines and their receptors closely associate with the temporal and spatial recruitment of neutrophils to wound sites during the inflammatory phase and promote the clearance of necrotic neutrophils during the transition into the proliferative phase. Functional aberrancy in these chemokines and chemokine receptor systems is recognized as one of the important mechanisms underlying the pathology of impaired wound healing. Future Directions: Future research should aim to investigate the therapeutic modulation of neutrophil activity through the targeting of specific chemokines or chemokine receptors in the early inflammatory phase to improve clinical management of wound healing.
    01/2014; DOI:10.1089/wound.2014.0559
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    ABSTRACT: Aberrant expression of CXCR4 in human breast cancer correlates with metastasis to tissues secreting CXCL12. To understand the mechanism by which CXCR4 mediates breast cancer metastasis, MCF-7 breast carcinoma cells were transduced to express wild-type CXCR4 (CXCR4WT) or constitutively active CXCR4 (CXCR4ΔCTD), and analyzed in two-dimensional cultures (2D), three-dimensional reconstituted basement membrane (3D rBM) cultures, and in mice using intravital imaging. Two-dimensional cultures of MCF-7 CXCR4ΔCTD cells, but not CXCR4WT, exhibited an epithelial to mesenchymal transition (EMT) characterized by up-regulation of ZEB-1, loss of E-cadherin, up-regulation of cadherin 11, p120 isoform switching, activation of ERK1/2 and MMP-2. In contrast to the 2D environment, MCF-7 CXCR4WT cells cultured in 3D rBM exhibited an EMT phenotype, accompanied by expression of CXCR2, CXCR7, CXCL1, CXCL8, CCL2, IL-6, and GM-CSF. Dual inhibition of CXCR2 with CXCR4, or inhibition of either receptor with inhibitors of MEK1 or PI3K, reversed the aggressive phenotype of MCF-7 CXCR4 expressing or MDA-MB-231 cells in 3D rBM. Intravital imaging of CXCR4 expressing MCF-7 cells revealed that tumor cells migrate toward blood vessels and metastasize to lymph nodes. Thus, CXCR4 can drive EMT along with an up-regulation of chemokine receptors and cytokines important in cell migration, lymphatic invasion and tumor metastasis.
    Molecular biology of the cell 01/2014; 25(5). DOI:10.1091/mbc.E13-07-0360 · 5.98 Impact Factor
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    ABSTRACT: Chiral nonracemic cis-4,5-bis(aryl)imidazolines have emerged as a powerful platform for the development of cancer chemotherapeutics, stimulated by the Hoffmann-La Roche discovery that Nutlin-3 can restore apoptosis in cells with wild-type p53. The lack of efficient methods for the enantioselective synthesis of cis-imidazolines, however, has limited their more general use. Our disclosure of the first enantioselective synthesis of (-)-Nutlin-3 provided a basis to prepare larger amounts of this tool used widely in cancer biology. Key to the decagram-scale synthesis described here was the discovery of a novel bis(amidine) organocatalyst that provides high enantioselectivity at warmer reaction temperature (-20 °C) and low catalyst loadings. Further refinements to the procedure led to the synthesis of (-)-Nutlin-3 in a 17 g batch and elimination of all but three chromatographic purifications.
    The Journal of Organic Chemistry 10/2013; 78(21). DOI:10.1021/jo401321a · 4.64 Impact Factor
  • Cancer Research 08/2013; 73(8 Supplement):2614-2614. DOI:10.1158/1538-7445.AM2013-2614 · 9.28 Impact Factor
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    ABSTRACT: Bone marrow-derived human mesenchymal stem cells (hMSCs) either promote or inhibit cancer progression, depending on factors that heretofore have been undefined. Here we have utilized extreme hypoxia (0.5% O2) and concurrent treatment with metal carcinogen (nickel) to evaluate the passage-dependent response of hMSCs toward cancerous transformation. Effects of hypoxia and nickel treatment on hMSC proliferation, apoptosis, gene and protein expression, replicative senescence, reactive oxygen species (ROS), redox mechanisms, and in vivo tumor growth were analyzed. The behavior of late passage hMSCs in a carcinogenic hypoxia environment follows a profile similar to that of transformed cancer cells (i.e., increased expression of oncogenic proteins, decreased expression of tumor suppressor protein, increased proliferation, decreased apoptosis, and aberrant redox mechanisms), but this effect was not observed in earlier passage control cells. These events resulted in accumulated intracellular ROS in vitro and excessive proliferation in vivo. We suggest a mechanism by which carcinogenic hypoxia modulates the activity of three critical transcription factors (c-MYC, p53, and HIF1), resulting in accumulated ROS and causing hMSCs to undergo cancer-like behavioral changes. This is the first study to utilize carcinogenic hypoxia as an environmentally relevant experimental model for studying the age-dependent cancerous transformation of hMSCs.-Crowder, S. W., Horton, L. W., Lee, H. H., McClain, C. M., Hawkins, O. E., Palmer, A. M. Bae, H., Richmond, A., Sung, H.-J. Passage-dependent cancerous transformation of human mesenchymal stem cells under carcinogenic hypoxia.
    The FASEB Journal 04/2013; 27(7). DOI:10.1096/fj.13-228288 · 5.48 Impact Factor
  • Cancer Research 02/2013; 73(3 Supplement):A53-A53. DOI:10.1158/1538-7445.TIM2013-A53 · 9.28 Impact Factor
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    ABSTRACT: Oncogene-induced senescence can provide a protective mechanism against tumour progression. However, production of cytokines and growth factors by senescent cells may contribute to tumour development. Thus, it is unclear whether induction of senescence represents a viable therapeutic approach. Here, using a mouse model with orthotopic implantation of metastatic melanoma tumours taken from 19 patients, we observed that targeting aurora kinases with MLN8054/MLN8237 impaired mitosis, induced senescence and markedly blocked proliferation in patient tumour implants. Importantly, when a subset of tumour-bearing mice were monitored for tumour progression after pausing MLN8054 treatment, 50% of the tumours did not progress over a 12-month period. Mechanistic analyses revealed that inhibition of aurora kinases induced polyploidy and the ATM/Chk2 DNA damage response, which mediated senescence and a NF-κB-related, senescence-associated secretory phenotype (SASP). Blockade of IKKβ/NF-κB led to reversal of MLN8237-induced senescence and SASP. Results demonstrate that removal of senescent tumour cells by infiltrating myeloid cells is crucial for inhibition of tumour re-growth. Altogether, these data demonstrate that induction of senescence, coupled with immune surveillance, can limit melanoma growth.
    EMBO Molecular Medicine 01/2013; 5(1). DOI:10.1002/emmm.201201378 · 8.25 Impact Factor
  • D. Raman, NL Duvall-Noelle, A. Richmond
    Cancer Research 12/2012; 72(24 Supplement):P1-05-01-P1-05-01. DOI:10.1158/0008-5472.SABCS12-P1-05-01 · 9.28 Impact Factor
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    Society for Melanoma Research Congress, 2012; 11/2012
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    ABSTRACT: The chemokine receptors, CXCR1 and CXCR2, couple to Gαi to induce leukocyte recruitment and activation at sites of inflammation. Upon activation by CXCL8, these receptors become phosphorylated, desensitized, and internalized. In this study, we investigated the role of different G protein-coupled receptor kinases (GRKs) in CXCR1- and CXCR2-mediated cellular functions. To that end, short hairpin RNA was used to inhibit GRK2, 3, 5, and 6 in RBL-2H3 cells stably expressing CXCR1 or CXCR2, and CXCL8-mediated receptor activation and regulation were assessed. Inhibition of GRK2 and GRK6 increased CXCR1 and CXCR2 resistance to phosphorylation, desensitization, and internalization, respectively, and enhanced CXCL8-induced phosphoinositide hydrolysis and exocytosis in vitro. GRK2 depletion diminished CXCR1-induced ERK1/2 phosphorylation but had no effect on CXCR2-induced ERK1/2 phosphorylation. GRK6 depletion had no significant effect on CXCR1 function. However, peritoneal neutrophils from mice deficient in GRK6 (GRK6(-/-)) displayed an increase in CXCR2-mediated G protein activation but in vitro exhibited a decrease in chemotaxis, receptor desensitization, and internalization relative to wild-type (GRK6(+/+)) cells. In contrast, neutrophil recruitment in vivo in GRK6(-/-) mice was increased in response to delivery of CXCL1 through the air pouch model. In a wound-closure assay, GRK6(-/-) mice showed enhanced myeloperoxidase activity, suggesting enhanced neutrophil recruitment, and faster wound closure compared with GRK6(+/+) animals. Taken together, the results indicate that CXCR1 and CXCR2 couple to distinct GRK isoforms to mediate and regulate inflammatory responses. CXCR1 predominantly couples to GRK2, whereas CXCR2 interacts with GRK6 to negatively regulate receptor sensitization and trafficking, thus affecting cell signaling and angiogenesis.
    The Journal of Immunology 08/2012; 189(6):2824-32. DOI:10.4049/jimmunol.1201114 · 5.36 Impact Factor
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    ABSTRACT: Although human angiosarcoma has been associated frequently with mutational inactivation of the tumor suppressor gene Ink4a/Arf, the underlying mechanisms have not been delineated. Here we report that malignant angiosarcoma is associated with high levels of RelA/NF-κB and IL-6 in contrast to normal vessels or benign hemagiomas. Studies of Ink4a/Arf deficient mice not only recapitulate genetic traits observed in human angiosarcoma, but also unveil a possible therapeutic link comprised of the NF-kB/IL-6/Stat3 signaling axis. In Ink4a/Arf(-/-) cells, NF-κB controlled Stat3 signaling by transcriptionally controlling the expression of IL-6, gp130, and Jak2. Further, IL-6 mediated Stat3 signaling through the sIL-6R. Inhibition of Ikkβ solely in myeloid cells was insufficient to block angiosarcoma development; in contrast, systemic inhibition of Ikkβ, IL-6, or Stat3 markedly inhibited angiosarcoma growth. Our findings offer clinical implications for targeting the NF-kB/IL-6/STAT3 pathway as a rational strategy to treat angiosarcoma. Cancer Res; 72(18); 4682-95. ©2012 AACR.
    Cancer Research 07/2012; 72(18):4682-95. DOI:10.1158/0008-5472.CAN-12-0440 · 9.28 Impact Factor

Publication Stats

5k Citations
722.04 Total Impact Points


  • 1993–2014
    • Vanderbilt University
      • • Department of Cancer Biology
      • • Department of Veterans Affairs
      • • Department of Molecular Physiology and Biophysics
      • • Department of Plastic Surgery
      • • Department of Medicine
      • • Vanderbilt Center for Stem Cell Biology
      Нашвилл, Michigan, United States
  • 2012
    • North Carolina Central University
      • Department of Biology
      Durham, NC, United States
  • 2002–2011
    • United States Department of Veterans Affairs
      Бедфорд, Massachusetts, United States
    • University of Louisville
      • Department of Pharmacology and Toxicology
      Louisville, Kentucky, United States
  • 2001–2009
    • U.S. Department of Veterans Affairs
      Washington, Washington, D.C., United States
  • 2007
    • Georgetown University
      Washington, Washington, D.C., United States
  • 2003–2005
    • Meharry Medical College
      Nashville, Tennessee, United States