Cell and Developmental Biology University of Michigan

Research

Development of immunotherapies for primary and metastatic brain cancer: from basic immunobiology mechanisms to translational immune-therapeutics. Tumor immune-microenvironment: its role in tumor progression and response to therapeutics. Cross talk between cancer cells and hematopoietic stem/progenitor cells. Mechanisms affecting the migration of immune cells from peripheral lymphoid organs to the tumor microenvironment

My research program covers the areas of cancer immunology, the tumor microenvironment, signaling via pattern recognition receptors, innate immunity, role of cancer derived ligands at modulating the maturation of the myeloid hematopoietic stem cell lineage, and the development of immune mediated and targeted gene therapeutic approaches for the treatment of cancers -both primary brain tumors (i.e., glioblastoma multiforme, GBM) and also metastatic brain cancer (e.g., melanoma, lung cancer, breast, colon and prostate). GBM is the most aggressive primary brain tumor; it has a five year survival rate of <5%. Attempts at eliciting a clinically relevant anti-GBM immune response in brain tumor patients have met with limited success, due to the brain's immune privileged status, tumor immune evasion, and a paucity of dendritic cells (DCs) within the central nervous system. Recently, we have uncovered a novel pathway for the activation of an effective anti-GBM immune response and therapy which involvesthe high-mobility-group box 1 (HMGB1) protein, an alarmin released from dying tumor cells, which acts as an endogenous ligand for Toll-like receptor 2 (TLR2) signaling on bone marrow-derived GBM-infiltrating DCs.

Using a combined immunotherapy/conditional cytotoxic approach that utilizes adenoviral vectors (Ad) expressing Fms-like tyrosine kinase 3 ligand (Flt3L) and Herpes Virus type 1 thymidine kinase (HSV1-TK) delivered into the tumor mass (Ali et al, Cancer Research, 2005), we demonstrated that cytotoxic T cells mediate both tumor regression and immunological memory. Infiltration of dendritic cells (DCs), and other immune cells, into the GBM microenvironment, clonal expansion of antitumor T cells, and induction of an effective anti-GBM immune response were TLR2 dependent. My group demontrated that the endogenous ligand responsible for TLR2 signaling on tumor-infiltrating DCs is HMGB1. Increased levels of HMGB1 were detected in the serum of tumor-bearing Ad-Flt3L/Ad-HSV1TK (+GCV)-treated mice. HMGB1 was also released from melanoma, small cell lung carcinoma, and glioma cells treated with radiation or temozolomide, indicating the universality of this mechanism. Administration of either glycyrrhizin (a specific HMGB1 inhibitor) or anti-HMGB1 immunoglobulins to tumor-bearing mice treated with Ad-Flt3L and Ad-TK, abolished therapeutic efficacy, demonstrating the critical role played by HMGB1-mediated TLR2 signaling to elicit tumor regression (Curtin et al, PLoS Medicine, 2009, 6 (1), e10). We are aiming to design and develop small molecules, HMGB1 agonists to be used as adjuvants for the treatment of solid cancers.

As an extension of these new data, my team has embarked on a research project which aims to uncover the molecular and cellular signaling cascades triggered by pattern recognition receptor ligands released by tumor cells. Our data indicates that these tumor derived ligands bind to pattern recognition receptors present on bone marrow derived hematopoietic stem cells (HSCs) and -as a consequence- they halt their maturation, thus inducing the accumulation of immature immunosuppressive cells, i.e., myeloid derived suppressor cells (MDSCs). We anticipate these studies will uncover previously unknown anti-cancer therapeutic targets, i.e., using small molecules and small interfering RNA technologies, aimed at modulating the maturation of MDSCs. These studies should also direct us to new research areas on the interphase between cancer and hematopoietic stem cell biology.

Lab Members

• Hikmat Assi
  Research Associate
• Alexandra Calinescu
  Post doctoral fellow
• Christopher Paran
  Research Technician
• Robert Doherty
  Research Technician
• Rosemary Lemons
  Lab Manager
• Sarah Suprise
  Undergraduate student
• Mohammed Saleh
  Undergraduate student
• Molly Dahlgren
  Administrative Assistant

Publications

Representative Publications

1. Sanderson NS, Puntel M, Kroeger KM, Bondale NS, Swerdlow M, Iranmanesh N, Yagita H, Ibrahim A,Castro MG, Lowenstein PR. (2012) Cytotoxic immunological synapses do not restrict the action of interferon-γ to antigenic target cells. Proc Natl Acad Sci U S A. 2012 May 15;109(20):7835-40.
2. Yohei Mineharu, Gwendalyn D. King, AKM G. Muhammad1, Serguei Bannykh, Kurt M. Kroeger, Chunyan Liu1, Pedro R. Lowenstein, and Maria G. Castro (2011) Engineering the Brain Tumor Microenvironment Enhances the Efficacy of Dendritic Cell Vaccination: Implications for Clinical Trial Design. Clin Cancer Res. 2011 Jul 15;17(14):4705-18. Epub Jun 1.
3. Candolfi M, Curtin JF, Yagiz K, Assi H, Wibowo MK, Alzadeh GE, Foulad D, Muhammad AK, Salehi S, Keech N, Puntel M, Liu C, Sanderson NR, Kroeger KM, Dunn R, Martins G, Lowenstein PR, Castro MG. (2011) B cells are critical to T-cell-mediated antitumor immunity induced by a combined immune-stimulatory/conditionally cytotoxic therapy for glioblastoma. Neoplasia Oct;13(10):947-60.
4. Candolfi M, Xiong W, Yagiz K, Liu C, Muhammad AK, Puntel M, Foulad D, Zadmehr A, Ahlzadeh GE, Kroeger KM, Tesarfreund M, Lee S, Debinski W, Sareen D, Svendsen CN, Rodriguez R, Lowenstein PR, Castro MG. (2010) Gene therapy-mediated delivery of targeted cytotoxins for glioma therapeutics. Proc Natl Acad Sci U S A. Nov 16;107
5. Larocque D, Sanderson NRS, Bergeron J, Curtin J, Girton J, Wibowo M, Bondale N, Kroeger KM, Yang J, Lacayo LM, Reyes KC, Farrokhi C, Pechnick RN, Castro MG, Lowenstein PR, (2010) Exogenous fms-like tyrosine kinase 3 ligand overrides brain immune privilege and facilitates recognition of a neo-antigen without causing autoimmune neuropathology. Proceedings of the National Academy of Sciences, USA. 107(32):14443-14448. Epub July 26
6. Candolfi M, Yagiz K, Foulad d, Ahlsadeh GE, Tesarfreund M, Muhammad AKM, Puntel M, Kroeger KM, Liu C, Lee S, Curtin J, King GD, Lerner J, Sato K, Lowenstein PR, Castro MG. (2009) Release of HMGB1 in response to proapoptotic glioma killing strategies: efficacy and neurotoxicity. Clinical Cancer Research 15(13):4401-4414. Portrayed on the Highlights Section. PMCID: PMC2769255
7. Curtin JF, Liu N, Candolfi M, Ziong W, Assi H, Yagiz Y, Edwards, MR, Michelsen KS, Kroeger KM, Liu C, Muhammad AKM, Clark M, Arditi M, Comin-Anduix B, Ribas A, Lowenstein PR,Castro MG. (2009) HMGB1 mediates endogenous TLR2 activation and brain tumor regression. PLoS Medicine Jan 13; 6(1):e10; PMCID: PMC2621261
8. Muhammad AKM, Candolfi M, King GD, Foulad D, Yagiz K, Mineharu Y, Kroeger K, Treuer KA, Nichols S, Sanderson NS, Lowenstein PR, Castro MG. (2009) Anti-glioma immunological memory in response to conditional cytotoxic/immune-stimulatory gene therapy: humoral and cellular immunity lead to tumor regression. Clinical Cancer Research 15(19):6113-6127, Epub 2009 Sep 29. PMCID: PMC2796687
9. King GD, Muhammad AKM G, Xiong W, Kroeger KM, Puntel M, Larocque D, Ng P, Lowenstein PR, Castro MG. (2008) High-capacity adenoviral vector-mediated anti-glioma gene therapy in the presence of systemic anti-adenovirus immunity. Journal of Virology 82(9):4680-4684; PMCID: PMC2293067
10. Curtin, JF, CandolfiM, Fakhouri TM, Liu C, Alden A, Edwards M, Lowenstein PR, Castro MG. (2008) Treg depletion inhibits efficacy of cancer immunotherapy: implications for clinical trials. PLoS ONE. Apr 23; 3(4):e1983; PMCID: PMC2291560
11. Curtin JF, King GD, Barcia C, Liu C, Hubert FX, Guillonneau C, Josien R, Anegon I, Lowenstein PR, Castro MG.(2006) Fms-like tyrosine kinase 3 ligand recruits plasmacytoid dendritic cells to the brain. Journal of Immunology 176:3566-3577. Portrayed on the cover.
12. Xiong W, Goverdhana S, Kroeger K, Ng P, Palmer D, Lowenstein PR, Castro MG, (2006) Regulatable Gutless Adenovirus Vectors Sustain InducibleTransgene Expression in the Brain in the Presence of an Immune Response Against Adenoviruses. Journal of Virology 80(1):27-37. Portrayed on the cover.
13. Ali S, King G, Curtin JF, Candolfi M, Xiong W, Liu C, Puntel M, Cheng Q, Prieto J, Ribas A, Kupiec-Weglinski J, van Rooijen N, Lassmann H, Lowenstein PR, Castro MG, (2005) Combined immuno-stimulation and conditional cytotoxic gene therapy provide long-term survival in a large glioma model. Cancer Research 65:7194-7204. Portrayed on the highlights section of Cancer Research.
14. Umana P, Gerdes CA, Stone D, Davis JR, Ward D, Castro MG, Lowenstein PR (2001) Efficient FLPe recombinase enables scalable production of helper-dependent adenoviral vectors with negligible helper-virus contamination. Nature Biotechnology, 19:582-585.
15. Thomas CE, Schiedner G, Kochanek S, Castro MG, Lowenstein PR (2000) Peripheral infection with adenovirus causes unexpected long-term brain inflammation in animals injected intracranially with first generation, but not with high-capacity adenovirus vectors: towards realistic long-term neurological gene therapy for chronic diseases. Proc. Natl. Acad. Sci. U.S.A., 97:7482-7487.
16. Dewey R, Morrissey G, Cowsill C, Stone D, Dodd NJF, Bolognani F, Southgate TD, Castro MG, Lowenstein PR, (1999) Chronic brain inflammation and persistent HSV1 TK expression in survivors of syngeneic glioma treated by adenovirus-mediated gene therapy. Nature Medicine 5,1256-63.