World Day Malaria Vector Biology and Control LIGHTNING TALKS Malaria Parasitology Breakout Room

All Presenters: Please be prepared to speak when your slide is queued up. Presentation order is indicated in the Lightning Talk Program circulated by email. The moderator will introduce you by name and affiliation, and then your time will start. To stay on time the moderator must advance to the next presentation at 3 minutes.

Attendees: This rapid- fire format does not allow for live questions, but we encourage you to contact any speaker for further networking. Identification of key determinants of cerebral malaria Sung-Jae Cha1, Xiang Yu2, Brian D. Gregory2, Tomoko Ishino3, Robert O. Opoka4, Chandy C. John5, and Marcelo Jacobs-Lorena1 [email protected] 1, Johns Hopkins School of , 2, University of Pennsylvania, 3, Ehime University, 4, Makerere University, 5, Indiana University 1. Cerebral Malaria (CM) 3. Plasma carries soluble CM-inhibitory factors Not all malaria patients develop CM.

Take home lesson: Liver vascular injury caused by sporozoite infection activates generation of CM inhibitory plasma factors. 2. Unexpected findings A B 4. Plasma marker assays identified sporozoite mediated liver injury activated blood stage anti-inflammatory & anti-apoptosis immune pathway including IGF-1.

5. IGF-1 6. Conservation with human suppresses CM Mortality

Silent exit Down stream of IGF-1 pathway is activated in CD68 WT CD68 KO CM-negative human and mouse CM vs No CM Development of a Rapid Diagnosis Test for Karli Redinger1, Quentin Watson2, Marlin Linger2, Rajeev Mehlotra2, Peter Zimmerman2, Jürgen Bosch1,3 1Division of Pediatric Pulmonology and Allergy/Immunology, Case Western Reserve University; 2Center for and Diseases, Case Western Reserve University; 3 InterRayBio, LLC Introduction Results

・There is currently no sensitive RDT that A B C specifically tests for Plasmodium vivax Malaria which requires additional treatment for eradication ・Five protein targets have been selected, expressed, and refolded using SAPN particles for the production of antibodies: PvEBP2, PvDBP, PvLDH, Figure 1. Antigen production and validation. A) Pvs25, and PvEBP2-H7 D E SAPN particles generated, protein and peptide- based B) Results are from a patient sample dilution ELISA assay probed with PvEBP2 antibodies. The red Aims line is the current detection limit for RDTs and the green line is our proposed detection limit based on ・Create a Rapid Diagnostic Test sensitive and this assay. C) Imagestream flow cytometry analysis D) Serum Samples from Malaria Endemic Regions specific for P.vivax using the target antibodies Ranked by Microscopy into P.vivax and P. falciparum Positives E) Limit of Detection of PvEBP2 generated in the lab Antibody. The signal for P.vivax is stronger than P.falciparum at all dilution concentrations. ・Determine affinity and specificity of antibodies towards P.vivax antigens compared to other Future Direction Contact Information Malaria species using Western blots, ELISA assays, ・ Conduct flow cytometry assays to determine Presenter: Karli Redinger ([email protected]) paper-based essays, and flow cytometry for double positive cell counts (infected + Ab) Primary Investigator: Jürgen Bosch ([email protected]) between Malaria species evaluation Artesunate response of isogenic lines differ in spleen-intact and splenectomized Aotus primates irrespective of PfK13 type Contact email: [email protected] Angela C. Ellisa , Shuchi N. Amina, John Gravesb, Surika Maharajb, Crystal Hiteb, Marvin L. Thomasb, Jianbing Mua, Juliana M. Sáa, and Thomas E. Wellemsa aLaboratory of Malaria and Vector Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA; bDivision of Veterinary Research, Office of Research Services, National Institutes of Health, Bethesda, MD 20892

Background Methodology Conclusion 1. P. falciparum recrudescences are common CRISPR-Cas9 gene editing (FVOK13C580→FVOK13C580Y) FVO K13 C580Y had after (ART) monotherapy higher RSAs than 2. Artemisinin-based Combination Therapies wildtype FVO (K13 C580) (ACT) are frontline antimalarial treatments 3. Parasite clearance times are highly dependent on the host immunity No significant differences in parasite clearance

half-life (t1/2) between isogenic clones in spleen- intact Aotus Collect clearance data at specified time-points • t1/2= 5.35 ± 0.76 hours Results for FVOK13 C580Y • t = 6.82 ± 0.79 hours Dondorp, A.M. et al, 2009, FVOK13C580 in A. l. griseimembra (Sp+) FVOK13C580Y in A. l. griseimembra (Sp+) 1/2 PMID: 19064625 2.0 2.0 AS-1 AS-1 AS-1 AI6016 AI5108 for FVO (K13 C580) AI5003 AS-1 AI5092 1.5 1.5 AS-1 AI6008 AI6014 1.0 1.0 Recrudescences 0.5 AS-1 0.5 Parasitemia (%) Parasitemia Parasitemia (%) Parasitemia happened irrespective of 0.0 0.0 0 10 20 30 40 50 0 10 20 30 40 50 Days post-inoculation Days post-inoculation PfK13 type and were not significantly different from each other

FVOK13C580Y in A. nancymaae (Sp-) FVOK13C580Y in A.nancymaae (Sp+) • Recrudescence: 11 ±

5.0 2.5 1963 AS-1 86244 4.5 AS-1 1.70 days for FVO 4.0 1964 2.0 86581 3.5 86431 86301 3.0 1.5 • Recrudescence: 7.63 ± 2.5 2.0 1.0 AS-1 K13 AS-1 1.5 AS-1 1.10 days for FVO

1.0 (%) Parasitemia 0.5 Parasitemia (%) Parasitemia C580Y 0.5 AS-1 0.0 0.0 0 10 20 30 40 0 10 20 30 40 Days post-inoculation Days post-inoculation *This work was supported by the NIAID

Fig 2. Parasite-clearance half-life (t1/2) plots. Plots show parasite clearance half-lives of FVO Division of Intramural Research Fig 1. Parasitemia development and clearance in Aotus primates. Plots show parasitemia isogenic pair in both spleen-intact and splenectomized Aotus animals. Plots obtained from the development, clearance, and recrudescences of parasites treated with AS (IV) at 4 mg/kg/day for 3 Worldwide Antimalarial Resistance Network (WWARN)’s Parasite Clearance Estimator (PCE). days. Sá, J.M. et al, 2018, PMID: 30455312 Machine Learning based models to Predict Synergism in Antimalarial Combinational Therapies against Plasmodium falciparum using a new Data Science Synergy Predictor tool (DS-SynPred) Angélica M. Rosado-Quiñones, María Del Mar Figueroa-Gispert, Jennifer Díaz-Rivera, Roberto G. Díaz-González, Kelvin Carrasquillo-Carrion, Roberto A. Feliu-Maldonado, Pedro Fernández-Gochez, Brenda G. Nieves-Rodríguez, Emilee E. Colón-Lorenzo, Abiel Roche-Lima, and Adelfa E. Serrano

Background Methodology

Antimalarial Resistance Data Science Synergy Predictor: A new computational tool to Combinational Therapy develop ML models for predicting synergistic Machine Learning combinations using antimalarial datasets of P. falciparum DS-SynPred tool strains.

Results

Conclusion DS-SynPred allows to develop ML models to predict synergism in combinatorial therapies for malaria and other diseases.

Funding: CCRHD NIH GRANT U54 MD007600 Do you have any questions? [email protected] EXP2 is important for intrahepatic development during Plasmodium liver stage Tahir Hussain, Jose Linera-Gonzalez, John M Beck, Manuel A Fierro, Gunnar R Mair, Ryan C Smith and Josh R Beck╫ Dept. of Biomedical Sciences, Iowa State University, Ames. IA hsp70 hsp70 Blood stage Liver stage EXP2 3X FLAG glmS nanoLUC EXP2 3X FLAG nanoLUC Quantitation of EXP2 knockdown

EXP2 UIS4 merge EXP2 UIS4 merge EXP2- FLAG- glmS EXP2- FLAG

intensity intensity

Glucosmaine (mM) Glucosmaine

1 0.5 0.3 UT 0.3 0.5 1

1 0.5 0.3 UT 0.3 0.5 1 EXP2 Mean Fluorescence Fluorescence Mean EXP2

Nanoluciferase assay shows significant EXP2 knockdown reduces merosome detachment Graphical summary decrease in parasite growth UT 0.3 0.5 1 ATQ

UT 0.3 0.5 1 ATQ glmS

glmS EXP2

control Control EXP2- FLAG- glmS EXP2- FLAG EXP2- FLAG- glmS EXP2- FLAG

EXP2

email ID: [email protected] Fluorescence in situ Hybridization (FISH) for Specific Identification of and . J. Shah1,2* and R. Ramasamy1 1ID-FISH Technology and 2 IGeneX, Milpitas, CA 95035, USA. [email protected]; [email protected]

FISH for Malaria Parasites 1. Whole blood treated with permeabilization buffer 2. Thin blood smear made & methanol fixed 3. Hybridization with probes (for 18S D rRNA) for 15 min at 370C 4. Washed and mounted slides fluorescence viewed in UV or LED microscope with filters 5. Genus probe – green & Species probe – red fluorescence 6. Detection limit = 55-80 parasites per μl blood E 7. Stable reagents; test time <<2h; detects only live cells; inexpensive A – P. falciparum B – P. knowlesi Shah et al. PLoS One. 2015;10(9):e0136726. for P. falciparum & P. vivax FISH C – P. vivax Shah et al. Parasites Vectors. 2017;10:342 for P. knowlesi FISH E D – P. malariae Shah et al. Diagnostics 2020;10:377 & 2020;10:761 for Babesia FISH E – P. ovale D World Malaria Day Malaria Vector Biology and Control Please rejoin the main meeting using your unique link.

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