Louisiana Research Day Program Book 2025

Biomedical Research: Section 2

Biomedical Research: Section 2

Md Towhidul Islam Tarun, MS 1 ; Heba E. Elsayed, PhD 1 ; Hassan Y. Ebrahim, PhD 1,2 ; Khalid A. El Sayed, PhD 1* 1 University of Louisiana Monroe, College of Pharmacy, Monroe, LA; 2 VCOM-Louisiana, Monroe, LA 27 THE OLIVE OIL PHENOLIC S-(-)-OLEOCANTHAL SUPPRESSES COLORECTAL CANCER PROGRESSION AND RECURRENCE BY MODULATING SMYD2-EZH2 AND C-MET ACTIVATION

Krishna Patel, BS; Sarah Voth, PhD 1 ; K. Adam Morrow, PhD 1 ; Rebekah L. Morrow, PhD 1 1 VCOM-Louisiana 28 THE ROLE OF AMYLOID PRECURSOR PROTEIN IN PULMONARY ENDOTHELIAL INFECTION

Context: Colorectal cancer (CRC) is the third most common cancer in the US and the second leading cancer-associated mortality cause. Available CRC therapies achieve modest outcomes and fail to prevent its recurrence. Epidemiological studies indicated that the Mediterranean diet rich in olive oil reduced CRC incidence. Objective: This study aimed at the identification and assessment of active anti-CRC olive phenolics. Methods: The MTT, wound healing, and colony formation assays were used to discover and assess the in vitro anti-CRC activity of olive phenolics. A nude mouse xenografting model was used to assess the in vivo CRC progression and recurrence suppressive activity of OC in pure and crude forms. OC isolated from olive oil using liquid-liquid extractions. Results: Screening of olive phenolics for in vitro antiproliferative activity against a diverse panel of CRC cell lines identified the extra-virgin olive oil (EVOO) S-(-)-oleocanthal (OC) as the most active hit. OC showed IC50 values of 4.2, 9.8,

14.5, and 4.9 μM against HCT116, COLO320 DM, WiDr, and SW48 CRC cells, respectively. The lysine methyltransferases SMYD2 and EZH2, along with the receptor tyrosine kinase c-MET proved aberrantly dysregulated in invasive and metastatic CRC. SMYD2 and c-MET were validated as OC molecular targets in multiple malignancies. Daily oral 10 mg/kg OC treatments over 15 days suppressed 72.5% of the KRAS mutant HCT-116-Luc cells tumor weight in male nude mice. Continued OC daily oral use after primary tumors surgical excision over additional 40 days significantly suppressed the HCT-116-Luc locoregional tumor recurrence and totally prevented the distant tumors recurrence. The SMYD2-EZH2 expressions and c-MET activation were notably suppressed by OC treatments in vitro and in collected animal primary tumors. Conclusions: OC and olive phenolics are potential nutraceutical interventions useful for the CRC control and prevention of its relapse.

Context: Toll-Like Receptors (TLRs) are pattern recognition receptors that act as part of the innate immune response in endothelial cells. Amyloidogenic proteins, such as β -amyloid can be employed as a defense mechanism against a variety of pathogens and may function with TLR pathways to aid in protection. The role of amyloid precursor protein in the setting of host-pathogen interaction in the pulmonary vasculature is unclear. Objective: The goal of this study was to determine whether amyloid precursor protein knock out (APPKO) enhanced sensitivity of pulmonary microvascular endothelial cells (PMVECs) to Pseudomonas aeruginosa infection. Methods: Whole cell lysates were collected, and RNA was isolated from wild type (MVR1D), CRISPR control (gRNA), and two separate APPKO cell lines (APPKO3/APPKO12). Whole cell lysates were subjected to immunoblot, and RNA was converted to cDNA prior to RT-PCR analysis to assess APP levels. All cell lines were infected with Pseudomonas strains Λ PcrV and ExoY to determine whether the presence

or absence of APP impacted bacterial growth and endothelial monolayer damage following inoculation. Bacterial plating assays were carried out to assess growth, and images were captured to assess monolayer integrity. Results: Western blots show decreased levels of APP in APPKO cells, while PCR revealed varying levels of message in APPKO cells. The absence of APP enhanced bacterial growth of both strains of Pseudomonas. APPKO cells show increased gap formation in response to both strains of Pseudomonas compared to control cells. Conclusions: We concluded that the absence of amyloid precursor protein enhanced sensitivity of pulmonary microvascular endothelial cells to Pseudomonas aeruginosa, potentially by allowing greater bacterial proliferation or survival.

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2025 Research Recognition Day