Professor Emeritus of Biological Sciences
Research Director of the Cancer Research Center
PhD, 1948 University Of Illinois - Urbana-Champaign
|Office:||Cancer Research Center|
Mutational patterns in aged Salmonella typhimurium: construction of strains for tumor destruction
Research descriptionAs bacterial cells age, an SOS signal invokes genomic changes, including point mutations, deletions, chromosomal translocations, large insertions, inversions, duplications and amplifications. These result from editing-defective-tRNA synthetases, genomic slip-strand-replication that inhibit repair of damaged DNA, and induction of gene-carrying phage that pop DNA in-and-out of genomes. Decades ago, Miloslav Demerec and associates had the foresight to isolate and catalog an extensive collection of thousands of Salmonella typhimurium auxotrophic mutants primarily for the purpose of inter- and intragenic chromosome mapping. Over 10,000 of the original sealed agar stab cultures, all stemming from a single parent, are now curated in our lab. The existence of these sealed, aged cultures offers a unique opportunity for mutational and evolutionary studies, especially since the complete genome of Salmonella typhimurium LT2 has been sequenced and annotated. Study of these mutants lead to the following hypotheses:  There is an association of observed mutations and chromosomal rearrangements with survival strategies, particularly those genetic differences that might have involved prophage/phage.  There is a co-evolution of S. typhimurium and associated phages. Phage genomic segments [either as prophage progenitors of lytic phage, as transducers of host genes, or as phage genome segments], upon recombination, have important roles in preservation and transfer of genes, including virulence factors.
Among the resulting progeny in our studies of mutation and survival strategies, we have selected a strain [CRC2631] that has lost all mammalian harmful effects, but has added affinity for tumor cells. To optimize these phenotypes we have engineered the strain to include  auxotrophy for thymine, aromatic amino acids, and histidine, and  deleted genes whose products might cause toxic shock. Together with Dr. Robert Kazmierczak, Dr. Heide Schatten, Alison Dino [laboratory manager], and several students, we have found that this strain has the unique ability to preferentially enter tumor cells, multiply in vacuoles, alter cytoskeletons and destroy mitochondria. Also, the lack of any toxic effect of this strain has been verified upon injection of over 10 million live bacterial cells into the peritoneal cavity of mice. In studies of the genes/gene products involved in the destruction of tumor cells, we have identified [with Richard Cummings, Emory U.] specific glycans involved in tumor destruction.
Kazmierczak, R.A., Best, A.A., Nguyen, D., Eisenstark, A. Whole-genome shotgun sequences of Salmonella enterica serovar Typhimurium Lilleengen type strains LT1, LT18, LT19, LT20, LT21, and LT22 (2017) Genome Announcements, 5 (30), art. no. e00720-17,
Kazmierczak, R.A., Gentry, B., Mumm, T., Schatten, H., Eisenstark, A. Salmonella bacterial monotherapy reduces autochthonous prostate tumor burden in the TRAMP mouse model (2016) PLoS ONE, 11 (8), art. no. e0160926.
Wang, C.-Z., Kazmierczak, R.A., Eisenstark, A. Strains, mechanism, and perspective: Salmonella -based cancer therapy (2016) International Journal of Microbiology, 2016, art. no. 5678702.
Kazmierczak, R., Choe, E., Sinclair, J., Eisenstark, A. Direct attachment of nanoparticle cargo to Salmonella typhimurium membranes designed for combination bacteriotherapy against tumors (2015) Methods in Molecular Biology, 1225, pp. 151-163.
Schatten, H., Eisenstark, A. Preface (2014) Salmonella: Methods and Protocols: Second Edition, pp. v-vi.
Kazmierczak, R., Choe, E., Sinclair, J., Eisenstark, A. Direct attachment of nanoparticle cargo to Salmonella typhimurium membranes designed for combination bacteriotherapy against tumors (2014) Salmonella: Methods and Protocols: Second Edition, pp. 151-163.
Choe, E., Kazmierczak, R.A., Eisenstark, A. Phenotypic evolution of therapeutic Salmonella enterica serovartyphimurium after invasion of TRAMP mouse prostate tumor (2014) mBio, 5 (4), art. no. e01182-14, 8 p.
Fagbohun, O.A., Kazmierczak, R.A., Petrenko, V.A., Eisenstark, A. Metastatic prostate cancer cell-specific phage-like particles as a targeted gene-delivery system (2013) Journal of Nanobiotechnology, 11 (1), art. no. 31.
Kazmierczak, R.A., Dino, A., Eisenstark, A., Schatten, H. New breast cancer treatment considerations: A brief review of the use of genetically modified (attenuated) bacteria as therapy for advanced and metastatic breast cancer (2013) Cell and Molecular Biology of Breast Cancer, pp. 363-371.
Chen, F., Liu, W.-Q., Liu, Z.-H., Zou, Q.-H., Wang, Y., Li, Y.-G., Zhou, J., Eisenstark, A., Johnston, R.N., Liu, G.-R., Yang, B.-F., Liu, S.-L. mutL as a genetic switch of bacterial mutability: Turned on or off through repeat copy number changes (2010) FEMS Microbiology Letters, 312 (2), pp. 126-132.
Eisenstark, A. Genetic Diversity among offspring from archived Salmonella enterica ssp. enterica serovar Typhimurium (Demerec Collection): In search of survival strategies (2010) Annual Review of Microbiology, 64, pp. 277-292.
Chen, F., Liu, W.-Q., Eisenstark, A., Johnston, R.N., Liu, G.-R., Liu, S.-L. Multiple genetic switches spontaneously modulating bacterial mutability (2010) BMC Evolutionary Biology, 10 (1), art. no. 277.