1. Title of Project/Research
Design and testing of bacteriocin peptides as an innovative, alternative therapeutic approach to
fight malaria.
Please submit a brief description of the scope of work of your research project, as well as
its objectives.
Resistance to antimalarial drugs has remained a large obstacle in the elimination of
malaria for decades, and has increasingly become a greater concern as drug resistance has been
reported in three of the five Plasmodium species that infect humans [1]. Moreover, reports
indicate that P. falciparum, the most virulent strain, has widespread resistance to almost every
class of antimalarial drugs, rendering current treatments largely ineffective [2]. Malaria remains
a global problem as 50% of the world’s population is at risk of infection, which results in 1
million deaths annually [2]. Currently, artemisinin-based combination therapies (ACTs) are the
most effective treatment in the fight against malaria [3]. Although this strategy has achieved
success, failure rates of ACTs have been increasing during the past decade due to the rise of
artemisinin-resistant parasites, primarily in Africa and Southeast Asia [4,5]. The susceptibility of
current treatment methods and the emergence of additional drug-resistant parasites will further
augment malaria cases and death, and greatly threaten the prior success of
elimination/eradication efforts. With the need for new drug alternatives, our goal is to design and
develop novel peptides as an alternative therapeutic means to treat and combat malaria.
Bacteriocins are a large class of natural antimicrobial peptides (AMPs) produced by
bacteria, which have been shown to exhibit antibiotic, anticancer, and antimalarial activities [6].
Peptides represent a newly emerging class of antimalarial agents, and several reports indicate
peptides exhibit activity against Plasmodium parasites in their blood and/or mosquito stages [6].
With short a half-life, good solubility, and membrane or metabolic disruption mechanisms,
bacteriocins make promising candidates as new antimalarial compounds over current drug
treatments. In order to enhance bacteriocin anti-Plasmodium activity, we describe a strategy to
design and screen bacteriocin peptides: 1. Using a human erythrocyte hemolysis assay, an initial
set of bacteriocin libraries will be screened for hemolytic activity to identify potentially toxic
peptides; 2. Bacteriocins will then be selected for antimalarial properties by using an in vitro
anti-Plasmodium activity assay to assess inhibition of asexual, blood-stage P. falciparum; 3. To
further optimize anti-parasite activity, bacteriocins found to exhibit activity will be analyzed for
important secondary structures as well as possible conserved motifs as scaffolds to iteratively
optimize the design of improved bacteriocin compounds. I anticipate that this strategy will lead
to the discovery of conserved, novel bacteriocin domains that can be further developed to
increase the effectiveness of the antimalarial activity of bacteriocin peptides.

2. References
1. Sinha, S., B. Medhi, and R. Sehgal. "Challenges of Drug-Resistant Malaria." Parasite
(Paris, France) 21 (2014): 61. Print.
2. Petersen, Ines, Richard Eastman, and Michael Lanzer. "Drug-Resistant Malaria:
Molecular Mechanisms and Implications for Public Health." FEBS letters 585.11 (2011):
1551-62. Print.
3. Egan, T. J., and C. H. Kaschula. "Strategies to Reverse Drug Resistance in
Malaria." Current Opinion in Infectious Diseases 20.6 (2007): 598-604. Print.
4. Enserink, M. "Malaria's Drug Miracle in Danger." Science (New York, N.Y.) 328.5980
(2010): 844-6. Print.
5. Vale, Nuno, Luísa Aguiar, and Paula Gomes. "Antimicrobial Peptides: A New Class of
Antimalarial Drugs?" Frontiers in Pharmacology 5 (2014) Print.
6. Aminake, M. N., et al. "Thiostrepton and Derivatives Exhibit Antimalarial and
Gametocytocidal Activity by Dually Targeting Parasite Proteasome and
Apicoplast." Antimicrobial Agents and Chemotherapy 55.4 (2011): 1338-48. Print.
7. Bell, Angus. "Antimalarial Peptides: The Long and the Short of it." Current
Pharmaceutical Design 17.25 (2011): 2719-31. Print.

3. Briefly describe your specific research role in your current project (250 words)
During my rotation in Dr. Shaun Lee’s laboratory, I performed the human erythrocyte
hemolysis assay on several bacteriocins from the Lee lab’s peptide library, and characterized the
ability of these peptides to lyse erythrocytes after a 24 hour exposure. I worked with Trupti
Pandharkar, a post-doc in Dr. Haldar’s laboratory, to screen a number of bacteriocins for anti-
Plasmodium activity using an in vitro assay. Throughout my rotation I also worked extensively
with Trupti to learn malaria culture techniques to maintain the 3D7 Plasmodium falciparum
asexual-stage parasites used in the assays. I performed a total of three anti-Plasmodium assays.
In two assays, the parasite culture was synchronized to ring or trophozoite/schizont stages to
assess how bacteriocins inhibit P. falciparum growth at different developmental stages. The third
assay used a non-synchronized culture which contained parasites at ring, trophozoite and
schizont stages. Although each of these assays were performed once, I obtained preliminary data
that suggests there are a number of candidate bacteriocins which exhibit antimalarial activity at a
micromolar concentration after 48 hours.

4. Please list or describe the broader societal impacts of your research. You may use bullet
points.
In 2012, there were approximately 700,000 reported deaths related to malaria worldwide
and 482,000 of these deaths occurred in children younger than five [1]. A majority of these
deaths were a result of infections by Plasmodium falciparum, which is reported to have clinical
resistance to many first-line antimalarial drugs [2]. Although malaria is distributed globally,
endemic areas are predominantly poor and developing countries within Africa and Southeast
Asia. With the emergence of new artemisinin-resistant parasites in underprivileged and largely
populated regions of the world, experts believe there is a possibility of a global outbreak of
malaria [3]. The established and emerging parasite resistance demands the production for new
and effective therapeutic compounds to treat malaria. Our antimalarial bacteriocins will serve as
novel therapeutics, and allow for safe and effective treatment to patients, while restraining the
emergence of drug-resistant parasites. Bacteriocins will also aid in reducing malaria transmission
by inhibiting merozoite Plasmodium growth within human hosts. In addition, malaria is an
economic burden to many nations worldwide. Malaria infection causes loss of short- and long-
term work productivity in infected individuals, amplifying poverty and limiting the growth of
endemic regions. The cost for treatment and prevention of malaria is also a significant weight on
personal and public expenses. The emergence and spread of drug resistance will only further
contribute to the financial burden as costs associated with malaria control will continue to rise.
However, the use of antimalarial bacteriocins to effectively combat Plasmodium parasites
provides an affordable and alternative method with widespread use to treat malaria.

5. References
1. Petersen, Ines, Richard Eastman, and Michael Lanzer. "Drug-Resistant Malaria:
Molecular Mechanisms and Implications for Public Health." FEBS letters 585.11 (2011):
1551-62. Print.
2. Tinto, H., et al. "In-Vitro Susceptibility of Plasmodium Falciparum to
Monodesethylamodiaquine, Dihydroartemisinin and Quinine in an Area of High
Chloroquine Resistance in Rwanda." Transactions of the Royal Society of Tropical
Medicine and Hygiene 100.6 (2006): 509-14. Print.
3. Sinha, S., B. Medhi, and R. Sehgal. "Challenges of Drug-Resistant Malaria." Parasite
(Paris, France) 21 (2014): 61. Print.