Kumi-Diaka, James

Genetic disorders like Cystic Fibrosis (CF) and X-linked Diseases (XLD) are inherited by offspring from parents who are healthy carriers of the autosomal recessive or allosomal genes. About 10-million Americans are healthy carriers of a mutant cysticfibrosis gene (predominantly F508del) and about 4% of newborns are at risk of being born with an X-linked disease. The current clinically approved mitigation plan for preventing genetic disorders in newborns from “at-risk couples” is to consider Preimplantation Genetic Testing for Monogenetic diseases (PGT-M). PGT-M involves an invasive microsurgical procedure that requires the removal of cells from 3-5day old embryos.
To minimize this invasiveness, we proposed a less invasive approach to prevent genetic disorders in newborns by genotypically sorting sperm cells which may be used for fertilization events (IUI/IVF/ICSI) with specially characterized antigens on the sperm surface membrane. For the disease models being adopted in our study – CF and XLD; we utilized certain monoclonal antibodies (mab) to target the H-Y male antigen and the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein which are both selectively expressed on the sperm surface.

Prostate cancer (PCa) is the second most diagnosed cancer in men. The resistance of prostate cancer to chemotherapy has been linked to the ATP Binding Cassette (ABC)-Mediated Multidrug Resistance (MDR). This study investigated the combination of 3-Bromopyruvate (3-BPA) and the anti-inflammatory molecule SC-514 in reducing MDR in prostate cancer. The compounds were incorporated into a PLGA nanoparticles to increase delivery to target cells.
To investigate the effectiveness of SC-514 and/3-BPA, cytoxicity assays including trypan blue dye exclusion, MTT tetrazolium reduction, NBT, LDH release poly caspase detection, cell titer glow assay, and ELISA were utilized. Both immunofluorescence and multidrug resistance efflux assays were utilized to estimate the number of drug resistant cells. SC-514 was encapsulated in PLGA nanoparticles via single-emulsion method. SC-514 nanoparticles were analyzed utilizing Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Liquid chromatography–mass spectrometry (LC–MS) was used to measure the amount of SC- 514 released from the nanoparticle. Alternative SC-514 drug release quantification methods such as colony forming assay, wound healing assay, and transwell and migration assay were explored.

The oncogenic role of many of inflammatory genes in prostate cancer (PCa) remains unexplored despite the increasing association of chronic inflammation with PCa initiation, progression, and therapy resistance. The overarching goal of this project was to identify dysregulated inflammatory genes that correlate with PCa progression and seek to understand their molecular mechanisms and the therapeutic potential of targeting them. To achieve this, we utilized cutting-edge integrative (epi) genomic and transcriptomic techniques to identify and characterize inflammatory genes whose deregulation or (epi) genetic alterations correlate with PCa progression.
Weighted Gene Co-expression Network Analysis and other multivariate analysis techniques identified IRAK1 as one of the inflammatory signatures found to be overexpressed in over 80% of prostate adenocarcinoma (PRAD) samples.
We also explored the diagnostic and prognostic potential of IRAK1 as a biomarker using Kaplan Meier Survival Analysis and AUROC Analysis. DNA methylation analysis showed that IRAK1 is hypomethylated and found to negatively correlate with its overexpression in PRAD patients. We also found some missense and truncated mutations in some patients and reported a high level of IRAK1 gene amplification in castration-resistant and neuroendocrine PCa patients.

Background: Prostate Cancer, in the absence of skin cancer, is the most prevalent type of cancer found
in the male population. Reactive Oxygen Species (ROS) can promote cancer cell proliferation when
they are at elevated levels. Vitamin C is a water-soluble antioxidant capable of inhibiting the formation
of ROS. Genistein, an isoflavone found in plants, also possesses the ability to inhibit ROS formation.
Objective To determine the potential therapeutic synergy between genistein and vitamin C and
investigate mechanism of action of genistein and/or vitamin C. Methods: Trypan blue assay was carried
out to know the % of viable cells. Varying concentrations of genistein with a constant concentration of
Vitamin C was used to treat LNCaP cells. After treatment of the cells with genistein and Vitamin C, MTT
assay of the cancer cells was performed and absorbance read through an ELISA reader. This gives the
values needed for interpreting cell viability after treatment. A statistical analysis performed to determine
whether the obtained results are statistically significant. Results: The results obtained from our
experiments are inconclusive with regards to the impact of Vitamin C on apoptotic cancer cell death
following genistein treatment. However the combination of genistein and vitamin C was more efficient in
tumor suppression than when the drugs were given separately. Conclusion: This study suggests that
treatment of prostate cancer using genistein can be enhanced by adjuvant treatment with vitamin C.
This study is of potential clinical success in reducing the cell death by necrosis.

In 2015, an estimated 220, 800 new cases and 27, 540 deaths are expected to occur due to prostate
cancer in US men, thus adding to the economic burden of the over 2.6 million men currently battling the
disease. Plethora of studies have demonstrated the phytotherapeutic potentials of beta-lapachone, a
phytochemical compound derived from the bark of the lapacho tree, native to South America. Betalapachone
(β-lap) has been shown to exhibit its anti-cancer effects majorly by the futile cycling between
the oxidized and the two electron reduction of β-lap mediated by NAD(P)H:quinone oxidoreductase
(NQO1) resulting in the generation of reactive oxygen species (ROS) using NADH or NAD(P) as
electron sources. β-lap is known to selectively kill human cancer cells, since NQO1 is expressed more
abundantly in numerous human solid tumors than in the adjacent normal tissues; NQO1 has been
shown to be exceptionally under expressed in hormone dependent prostate cancer cells (LNCaP)
compared to the hormone independent prostate cancer cells (PC3). This study was aimed to
investigate the enhancing effects of very low dose radiation (VLDR (20mGy)) derived from a
pyroelectric crystal generator on the phytotherapeutic activity of beta-lapachone in LNCaP cell line in
vitro accessed by MTT and Trypan blue assay. Treatment-induced intracellular levels of ROS were also
assessed using Nitro blue tetrazolium assay. NQO1 activities in LNCaP cells were also investigated
following treatment with VLDR and/or β-lap using Dicoumarol (NQO1 inhibitor). Results indicate that
LNCaP cells respond significantly to combined treatments compared to single treatments.

Prostate cancer after many years is still the second most common cancer in American
men with about 233,000 new cases and 29,480 deaths estimated to be occurring in 2014.
Despite the wide spectra of reports demonstrating the anti-cancer phytotherapeutic potentials of
beta-lapachone and soybean-derived genistein in various tumors, little emphasis had been placed
on their synergistic effects in androgen-independent PC3 and androgen-dependent LNCaP
prostate cancer cell lines. In this study, we aim to characterize the combined effects of genistein
and b-lapachone on the phyto/chemosensitivity of LNCaP and PC3 human prostate cancer cells
in-vitro, using MTT assay and LDH assay to study treatment-induced growth inhibition and
cytotoxicity. Annexin-V-FITC and PI-TUNEL assays were also used to determine the potential
treatment-induced apoptosis and/or necrosis.
Our results revealed that both PC3 and LNCaP are phytosensitive to both single and combined
treatments, though time-and dose-dependent. We observed that our treatments induced dual
death pathways-apoptosis and necrosis-in both cell types and also observed that growth
inhibition in both correlated positively with cell death in which, b-lapachone and genistein
induced cell cycle arrest at the G1 and/or S phase and G2–M checkpoints respectively.
Invariably, our results indicate that combination treatments with b-lapachone and genistein are
more potent in killing both PC3 and LNCaP cancer cells than treatment with either genistein or
b-lapachone alone. Our current results are therefore in agreement with the hypothesis that drugcombinations
that target cell cycles at different critical checkpoints are more effective in causing
cell death.

In spite the heavy investments in therapeutic research breast cancer still impacts the
lives of women globally. The projected incidence of new cases in USA for 2008 is 67,770,
with estimated 40,480 deaths. In this study, we investigated the therapeutic efficacy of
Cytoreg®-genistein combination treatment on MCF-7 human breast cancer cells. MCF-7
cells were treated with genistein and Cytoreg® single and combination treatments for 24-
48hr; and the chemosensitivity assessed using bioassays: Trypan Blue and MTT for cell
viability; Ethidium bromide/Rhodamine 123 to assess apoptosis induction; F AM PolyCaspase
binding assay for mechanism of action. The overall data indicated dose- and timedependent
cell death in the MCF-cells and that apoptosis was the major means of treatmentinduced
growth inhibition. There was evidence of Cytoreg®-induced autophagy in the cells.
The overall findings indicated that genistein-Cytoreg® combination was more efficacious
than either genistein or Cytoreg® alone. Cytoreg® enhanced the phytosensitivity of MCF-7
cells to genistein isoflavone.

This study determined the impact of vitamin C dose on genistein-induced apoptosis in LNCaP cancer cells at various treatment regimens in vitro. Although the linear regression of viability assay (MTT) indicated a p-value = 0.11; NBT assay reveal a declining SOD activity during cell death. Apoptosis induction was the main mode of treatment induced cell death. The overall data showed the trend of treatment efficacy as;(Gen 10uM + Vit C 40uM) > (Gen 30uM + Vit C 40uM) > (Gen 70uM + Vit C 40uM) > 10uM genistein > 70uM genistein. The chi-square test for comparing necrosis, apoptosis and life cells showed that Vitamin C could impact genistein-induced apoptosis in LNCaP cells (p = 0.0003). This study forms the basis for in vivo studies of the impact of vitamin C on genistein-induced apoptosis in LNCaP prostate cancer cells.

An estimated 220,800 new prostate cancer cases and 27,540 deaths are expected to occur in US men by the end of 2015. Despite the increased treatment modes for prostate cancer, there is still no definite cure, and prognosis remains, at best, cautiously optimistic. The explicit amalgamation of two or more cancer therapeutic modalities such as surgery, radiation, and chemotherapy, has been one of the main interests of clinical investigation for several decades. Genistein (GN) and Beta-lapachone (BL) are two of the most promising anticancer phytochemical compounds. However, the anticancer activities of BL have been correlated with the enzyme activity of NQO1. The aim of this study was to investigate the enhancing effects of VLDR derived from a portable pyroelectric crystal generator on the chemopreventive and/or chemotherapeutic effects of GN and BL in NQO1+ PC3 and NQO1± (deficient) LNCaP prostate cancer cells (PCa) in vitro. The combination treat ment-induced cytotoxicity was investigated via MTT and Trypan blue exclusion assays. Dicoumarol (an NQO1 inhibitor) was co-administered to assess the effect of VLDR on NQO1 modulation. Nitro-blue tetrazolium assay was used to assess the intracellular ROS levels. Fluorescence microscopy was also used to assess the mode of cell death. In this study, a novel quantitative modeling approach was employed to comparably assess the cytotoxic effects of specific drugs used alone or in combinations with VLDR and to predict the potential synergistic therapeutic combinations. The data suggests that VLDR induced a rise in ROS levels, followed by upregulation in NQO1 levels. Pharmacodynamic indices were developed to quantify and characterize the combination treatment as synergistic, additive or antagonistic per dose or time-interval. Synergism was found to be dose and time-interval dependent. The major mode of cell death by this combination therapeutic regimen was found to be via apoptosis . In conclusion, our results confirm that VLDR enhanced cytotoxicity effects of both drugs dose- and time-dependently.

Prostate cancer is one of the leading causes of death in men aged 40-55. Genistein isoflavone (4', 5', 7-trihydroxyisoflavone) is a dietary phytochemical with demonstrated anti-tumor activities in a variety of cancers. Topotecan Hydrochloride (Hycamtin) is an FDA-approved chemotherapy drug, primarily used for secondary treatment of ovarian,cervical and small cell lung cancers. This study was to demonstrate the potential anticancer activities and synergy of topotecan-genistein combination in LNCaP prostate cancer cells. The potential efficacy and mechanism of topotecan/genistein-induced cell death was investigated... Results: The overall data indicated that i) both genistein and topotecan induce cellular death in LNCaP cells, ii) topotecan-genistein combination was significantly more efficacious in reducing LNCaP cell viabiligy compared to either genistein or topotecan alone, iii) in all cases, cell death was primarily through apoptosis, via the activation of the intrinsic pathway, iv) ROS levels were increased and VEGF expression was diminished significantly with the topotecan-genistein combination treatment, v) genetic analysis of topotecan-genistein treatment groups showed changes in genetic expression levels in pathway specific apoptotic genes.... Conclusion: Treatments involving topotecan-genistein combination may prove to be an attractive alternative phytotherapy of adjuvant therapy for prostate cancer.