Combating Breast Cancer with the Wellderly Immune Repertoire
|Institution:||Scripps Research Institute|
Brunhilde Felding , Ph.D. -
|Award Cycle:||2011 (Cycle 17)||Grant #: 17NB-0058||Award: $473,750|
|Award Type:||IDEA Competitive Renewal|
|Detection, Prognosis and Treatment>Innovative Treatment Modalities: search for a cure|
Initial Award Abstract (2011)
Note: this project is a continuation of an earlier IDEA grant to the PI funded by the CBCRP from 2009-2010.
The “Wellderly” is a group of 1000 healthy people in their eighties who never had cancer or any other chronic disease. Our idea is that many of these people have a strong immune system, and that some may have had developing breast cancer cells in their bodies, at some point in time, but successfully produced antibodies that prevented tumor establishment. Our project will tap into the genes of the “Wellderly” by using a “library” from their immune cells that we generated. The library will be used to fish out antibodies that can inhibit breast cancer cells, particularly those that cause inflammatory breast cancer, the most aggressive form of this disease.
For these studies we established a unique new model of triple-negative, inflammatory breast cancer that reflects progression of a particularly treatment-resistant tumor type seen in the clinic. This model comprises cell lines from the primary tumor, local recurrence and pleural effusion that a young patient developed despite extensive regimens of chemotherapy and radiation. We are studying the immune repertoire of the Wellderly to detect antibodies that can block breast cancer growth and progression to an invasive phenotype. We are further performing genetic analyses of the increasingly aggressive and therapy-resistant tumor cells in our model to understand the underlying molecular basis related to breast cancer metastasis.
In more technical terms, we will: (1) use a combinatorial phage display scFv antibody library that we generated from the “Wellderly”, to identify new antigens expressed by the increasingly aggressive breast cancer cells during their epithelial-to-mesenchymal transition (EMT) which we established as a disease driving pathway in our breast cancer progression model; and (2) utilize a high-throughput screening platform for identifying growth inhibitory and EMT reversing scFvs. We have expanded the genetic analyses of our progression model and are beginning to understand gains and losses of chromosomal regions in relation to gene expression regulation. Our comparisons of the aCGH (comparative genome hybridization) and gene expression data, as well as functional network analyses will enable us to generate a synopsis for understanding gene regulation in the progression of a very aggressive and treatment resistant form of breast cancer.
Our hope is that Information on antigens against which growth inhibitory and EMT reversing antibodies are directed, and results from gene expression and chromosomal aberration analyses of our model, will identify novel therapeutic targets in aggressive breast cancer. Human antibodies from the “Wellderly” might be directly used in patients, or could be optimized for efficient therapy and combination treatments. Future study on if and how results from this work relate to triple negative, basal type breast cancers in general, may help find treatments for very aggressive and rapidly progressing types of breast cancer.
Final Report (2014)
Information and tools generated in this study lead to a better understanding of mechanisms of breast cancer progression in a particularly aggressive type of the disease. Results from this project not only help to open new avenues for exploring as lead mechanisms of breast cancer progression, but they also provide the research community, and potentially the clinical medical community with tools to target key molecular features of an aggressive breast cancer phenotype. Our genomic analyses of gene expression patterns manifested in a patient derived new and unique progression model of metaplastic, triple negative and inflammatory breast cancer revealed several key pathways as driving forces of disease progression in this highly aggressive breast cancer. Among these key drivers are clear alterations in cytoskeletal proteins and proteases – prominently including matrix degrading metallo proteinases - all of which indicate acquisition of a highly motile and invasive phenotype during disease progression. Furthermore, collective loss of cell-cell adhesion molecules clearly indicates loss of epithelial integrity and ability to disperse out of a tissue collective. Within this group of alterations, this cell model provided unique new clinical indication of epithelial to mesenchymal transition (EMT) within the short period of primary tumor formation, local – therapy resistant recurrence – and ultimately metastatic distant dissemination. Among the changes were also a unique new set of chemokines, some of which are novel and have never been associated with breast cancer previously.
In a separate group of changes, we found loss of tumor suppressor gene expression, prominently including PTEN, already at the level of primary tumor development. However, we also discovered loss of less explored genes with putative tumor suppressor activity. Failure of the progression cells from the local, treatment resistant recurrence and distant metastasis to express these genes was found to be caused by focal and/or interstitial deletion at the chromosomal level. The genomic analyses and functional studies carried out to validate expression alterations and functional contributions of altered gene products to breast cancer progression have spurred several new directions of research in our laboratory. Thus, this project has been seminal for a number of key discoveries of new pathways and enrichment of knowledge in already reported pathways found to be involved in breast cancer development and malignant progression. Panning of our Wellderly immune repertoire antibody library revealed unique new functional targets in progressing breast cancer: 1. Apolipoprotein E, ApoE, the major apoprotein of the chylomicron and ligand of distinct cell surface receptors involved with lipid uptake and lipid induced cells signaling pathways. 2. LRP8, one of the major cell surface receptors for AopE that is involved with control of cell migration and invasion.
Thus, the information and tools from this study provide novel, unexpected leads as a basis for development of improved and effective therapies against breast cancer progression.
Site-specific coupling and setrically controlled formation of multimeric antibody fab fragments with unnatural amino acids.
Periodical:Journal of Molecular Biology
Index Medicus: J Mol Biol
Authors: Hutchins BM, Kazane SA, Staflin K, Forsyth JS, Felding-Habermann B, Schultz PG, Smider VV
|Yr: 2011||Vol: 406||Nbr: 4||Abs:||Pg:595-603|
Contribution of platelets to tumour metastasis.
Periodical:Nature Reviews. Cancer
Index Medicus: Nat Rev Cancer
Authors: Gay LJ, Felding-Habermann B.
|Yr: 2011||Vol: 11||Nbr: 2||Abs:||Pg:123-34|
Comparison of in vitro and in vivo approaches to studying brain colonization by breast cancer cells.
Periodical:Journal of Neurooncology
Index Medicus: J. Neoroonc.
Authors: Loger M, Lee H, Forsyth JS, Felding-Habermann B
|Yr: 2011||Vol: 104||Nbr: 3||Abs:||Pg:689-696|
Selective formation of covalent protein heterodimers with unnatural amino acids.
Periodical:Chemistry and Biology
Index Medicus: Chem Biol
Authors: Hutchins BM, Kazane SA, Staflin K, Forsyth JS, Felding-Habermann B, Smider VV, Schultz PG
|Yr: 2011||Vol: 18||Nbr: 3||Abs:||Pg:299-303|
Chemically programmed antibodies targeting multiple av integrins and their effects on tumor-related functions in vitro.
Index Medicus: Bioconjug Chem
Authors: Goswami RK, Bajjuri KM, Forsyth JS, Das S, Hassenpflug W, et al.
|Yr: 2011||Vol: 22||Nbr: 8||Abs:||Pg:1535-1544|
Platelets alter tumro cell attributes to propel metastasis: programming in transit.
Authors: Gay LJ, Felding-Habermann B.
|Yr: 2011||Vol: 20||Nbr: 5||Abs:||Pg:553-554|
Synthesis of site-specific antibody-drug conjugates using unnatural amino acids.
Periodical:Proceedings of the National Academy of Sciences of the United States of America
Index Medicus: Proc Nat Acad Sci, U S A
Authors: Jun Y, Axup JY, Bajjuri KM, Ritland M.et al.
|Yr: 2012||Vol: 109||Nbr: 40||Abs:||Pg:16101-6|
Mitochondrial complex I activity and NAD+/NADH balance regulate breast cancer progression.
Periodical:Journal of Clinical Investigation
Index Medicus: J Clin Invest
Authors: Santidrian AF, Matsuno-Yagi A., Rittland M., Seo BB., Leboeuf SE, Gay LJ, et al.
|Yr: 2013||Vol: 123||Nbr: 3||Abs:||Pg:1068-1081|