Noor A. Hussein

All Publications

• Using mass cytometry to probe the STAT signaling landscape in circulating immune cells in Rheumatoid Arthritis uncovers signaling dysregulation and correlation with disease activity. Frontiers in medicine Macaubas, C., Bayram, B., Hussein, N., Jager, A., Davis, K. L., Graf, J., Nakamura, M., Zisman, D., Mellins, E. D. 2025; 12: 1622537

  Abstract

  The phosphorylation state of signaling proteins is a useful measure of cell function. The high dimensionality of mass cytometry (CyTOF) allows assessment of a large number of parameters using limited amount of material. By combining CyTOF high dimensionality with fixation of blood cells close to collection, cell phosphorylation patterns are preserved as close as possible to in vivo conditions. We used Cytof and whole fixed blood to investigate the phosphorylated forms of five Signal Transducers and Activators of Transcription - STATs - in Rheumatoid Arthritis (RA) patients and controls.We designed an antibody panel identifying 34 immune subpopulations combined with the phosphorylated (p) forms of Stat1, Stat3, Stat4, Stat5 and Stat6. Whole fixed blood from 21 RA patients and 10 healthy controls were analyzed by CyTOF. The frequency of immune cell subpopulations and the level of the pSTATs proteins were compared between the samples from RA patients and controls using Multiple Mann Whitney tests and clustering analysis. Correlation of the tested parameters with DAS28-ESR, DAS28-CRP and CDAI was examined.Levels of pSTAT5 were elevated in several CD4 T cell subsets, and positively correlated with DAS28-ESR and DAS28-CRP. pSTAT1 levels were elevated in CD4 T cell subpopulations, as well as in some subsets of CD8 T cell subsets, NK cells, monocytes and Dendritic cells. Levels of pSTAT6 in several CD4 T cell subpopulations, including T regulatory cells, were lower in samples from RA patients compared to controls, and negative correlations between CDAI and pSTAT6 were found. The frequency of CD4 T cell effector memory cells expressing the chemokine receptors CCR2 and CCR5 was lower in samples from RA patients compared to controls.We found distinct pattern of STAT proteins phosphorylation associated with circulating immune cells in RA samples compared to healthy controls. These findings correlated with measures of disease activity. Analysis of phosphorylation patterns may be useful to understand disease pathology, and also as a marker of treatment response and disease outcome. By avoiding cell isolation, phosphorylation patterns in several cell subpopulation can be analyzed close to the in vivo cell profile.

  View details for DOI 10.3389/fmed.2025.1622537

  View details for PubMedID 41446843

  View details for PubMedCentralID PMC12722927

• Sex-associated and disease state-dependent monocyte polarization and CNS-trafficking phenotypes in pediatric acute-onset neuropsychiatric syndrome (PANS). Journal of neuroinflammation Rahman, S. S., Hussein, N., Galfrè, S. G., Gaertner, F., Macaubas, C., Chan, A., Columbo, L., Gao, J., Galehdari, S., Bayram, B., Ma, M., Manko, C., Miles, K., Farhadian, B., Silverman, M., Thienemann, M., Or-Geva, N., Van Haren, K., Nadeau, K. C., Tian, L., Frankovich, J., Mellins, E. D. 2025; 22 (1): 273

  Abstract

  Pediatric acute-onset neuropsychiatric syndrome (PANS) is characterized by the sudden onset of obsessive-compulsive symptoms alongside a constellation of neuropsychiatric and somatic features. Disease progression typically includes flare and recovery states, with some patients exhibiting a persistent disease course (> 12 months of flare). We characterized circulating monocyte subsets during flare and recovery in pediatric patients with PANS, uncovering disease-state-dependent shifts in polarization and trafficking phenotypes. Inflammatory M1-like monocytes and monocyte-derived dendritic cells were elevated during flare, while anti-inflammatory M2-like monocytes were enriched in recovery. We also identified a circulating subset with a surface phenotype consistent with central nervous system (CNS) homing, which was reduced during flare and restored in recovery. These cells were detectable in the cerebrospinal fluid (CSF) of new-onset patients but not in persistent cases, suggesting differential compartmentalization during disease progression. Notably, monocyte phenotypes, including M2 polarization (monocytosis) and circulating CNS-homing profiles, exhibited striking sex-associated differences, particularly during recovery, with several subsets enriched in males but not females. Plasma from flare-phase patients modestly induced CNS-homing markers in monocytes from healthy donors, indicating the presence of circulating modulators. Together, these findings support a model in which distinct myeloid phenotypes-including sex-biased patterns-may contribute to both the pathogenesis and resolution of neuroinflammation in PANS.

  View details for DOI 10.1186/s12974-025-03549-6

  View details for PubMedID 41254741

  View details for PubMedCentralID 4340335

• Revisiting strategies to target ABC transporter-mediated drug resistance in CNS cancer. Cancer biology & medicine Amawi, H., Hammad, A. M., Hall, F. S., Hussein, N., Rataan, A. O., Mrayyan, A., Al-Kofahi, T., Hmedat, A., Ashby, C. R., Tiwari, A. K. 2025

  Abstract

  A significant number of anticancer drugs fail to treat primary and metastatic brain tumors primarily because of the complex blood-brain barrier (BBB) and overexpression of ATP-binding cassette (ABC) transporters, which decrease drug penetration into the central nervous system and ultimately into tumors. It is noteworthy that the ABC transporters, ABCB1 [known as P-glycoprotein (P-gp)] and ABCG2 [known as breast cancer resistance protein (BCRP)], are overexpressed in brain tumors, including common gliomas. The co-presence of these transporters may negate the inhibition of either transporter, particularly if both transport the same anticancer drug. The cellular export of drugs by ABC transporters has been implicated in mediating resistance to anticancer drugs. However, the clinical relevance as a therapeutic target in human tumors remains a matter of contention. Although effective and clinically approved ABC transporter inhibitors could potentially overcome drug resistance, none are currently approved. Furthermore, the ABC transporter inhibitors in clinical trials produced low or no clinical efficacy, significant toxicities, and unsuitable pharmacokinetic profiles. Therefore, innovative approaches are needed to efficaciously and simultaneously inhibit these transporters to surmount anticancer drug resistance. This review emphasizes the clinical significance of ABC transporters in diminishing the efficacy of brain tumor treatments. The molecular alterations in BBB following brain tumor development, which are linked to various cancer therapies, are discussed. The overexpression of ABCB1 and ABCG2 at the BBB is discussed, potential strategies to decrease the export of chemotherapeutics by these transporters and the associated challenges and failures are discussed, and the implementation of novel approaches is considered.

  View details for DOI 10.20892/j.issn.2095-3941.2025.0060

  View details for PubMedID 41024626

• Mitochondria as Regulators of Nonapoptotic Cell Death in Cancer. MedComm Malla, S., Neupane, R., Sood, S., Hussein, N., Abou-Dahech, M., Terrero, D., Ashby, C. R., Babu, R. J., Tiwari, A. K. 2025; 6 (8): e70244

  Abstract

  Mitochondria are involved in cell survival and metabolic processes including adenosine triphosphate production, heme biosynthesis, reactive oxygen species, and iron and calcium homeostasis. Although mitochondria are well known to contribute to apoptosis, a growing body of evidence indicates that mitochondria modulate nonapoptotic cell death (NACD) mechanisms, including autophagy, necroptosis, ferroptosis, paraptosis, pyroptosis, parthanatosis, and cuproptosis. These NACD pathways differ in molecular triggers, morphological characteristics, and immunological consequences, but they all involve mitochondria. For example, mitochondrial ROS and lipid peroxidation play a role in ferroptosis, whereas mitochondrial depolarization and the release of apoptosis inducing factor are paramount to parthanatosis. Mitochondrial swelling is a hallmark of paraptosis, whereas mitochondrial disruption is associated with pyroptosis. Autophagy, though primarily a survival mechanism, is also regulated by mitochondrial dynamics in cancer cells. In cuproptosis, mitochondrial protein aggregates when iron-sulfur cluster proteins are disrupted, resulting in copper-dependent cell death. There are many factors that influence NACD, including mitochondrial membrane potential, bioenergetics, calcium flux, metabolites, and interactions with the endoplasmic reticulum. The review comprehensively summarizes our understanding of mitochondrial and NACD interactions, particularly in cells resistant to classical apoptosis agents. Therapeutic vulnerabilities associated with mitochondria-mediated NACD could lead to next-generation therapies.

  View details for DOI 10.1002/mco2.70244

  View details for PubMedID 40703196

  View details for PubMedCentralID PMC12284444

• A SARS-CoV-2 vaccine on an NIR-II/SWIR emitting nanoparticle platform. Science advances Jiang, Y., Sanyal, M., Hussein, N. A., Baghdasaryan, A., Zhang, M., Wang, F., Ren, F., Li, J., Zhu, G., Meng, Y., Adamska, J. Z., Mellins, E., Dai, H. 2025; 11 (6): eadp5539

  Abstract

  The COVID-19 pandemic caused a global health crisis that resulted in millions of deaths. Effective vaccines have played central roles in curtailing the pandemic. Here, we developed a down-converting near-infrared IIb (NIR-IIb; 1500 to 1700 nanometers) luminescent, pure NaErF4@NaYF4 rare-earth nanoparticle (pEr) as vaccine carriers. The pEr nanoparticles were coated with three layers of cross-linked biocompatible polymers (pEr-P3; ~55 nanometers) and conjugated to the receptor binding domain (RBD) of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein. Upon subcutaneous injection of the pEr-P3-RBD nanovaccine in mice, in vivo NIR-IIb imaging revealed active vaccine trafficking and migration to lymph nodes through lymphatic vessels. Two doses of the adjuvant-free vaccine elicited long-lasting (>7 months) high titers of serum viral neutralization antibody and anti-RBD immunoglobulin G, along with robust RBD-specific germinal center B cells and T follicular helper cells. We devised in vivo NIR-II molecular imaging of RBD-specific cells in lymph nodes, opening noninvasive assessments of vaccine-elicited immune responses longitudinally.

  View details for DOI 10.1126/sciadv.adp5539

  View details for PubMedID 39919189

• Safety Results from a Phase 1 Double-blind Randomized Clinical Trial of Allogeneic Mesenchymal Stem Cells in Early RA Singer, N., Breitman, M., Haghiac, M., Gordesky, L., Reese, J., Lewis, S., Barnboym, E., Hussein, N., Lasalvia, S., Mellins, E., Bonfield, T., Anthony, D., Caplan, A., Lazarus, H. WILEY. 2023: 2575-2576

  View details for Web of Science ID 001190014302314

• A Novel Dialkylamino-Functionalized Chalcone, DML6, Inhibits Cervical Cancer Cell Proliferation, In Vitro, via Induction of Oxidative Stress, Intrinsic Apoptosis and Mitotic Catastrophe. Molecules (Basel, Switzerland) Len, J. M., Hussein, N., Malla, S., Mcintosh, K., Patidar, R., Elangovan, M., Chandrabose, K., Moorthy, N. S., Pandey, M., Raman, D., Trivedi, P., Tiwari, A. K. 2021; 26 (14)

  Abstract

  In this study, we designed, synthesized and evaluated, in vitro, novel chalcone analogs containing dialkylamino pharmacophores in the cervical cancer cell line, OV2008. The compound, DML6 was selective and significantly decreased the proliferation of OV2008 and HeLa cells in sub-micromolar concentrations, compared to prostate, lung, colon, breast or human embryonic kidney cell line (HEK293). DML6, at 5 μM, arrested the OV2008 cells in the G2 phase. Furthermore, DML6, at 5 μM, increased the levels of reactive oxygen species and induced a collapse in the mitochondrial membrane potential, compared to OV2008 cells incubated with a vehicle. DML6, at 5 μM, induced intrinsic apoptosis by significantly (1) increasing the levels of the pro-apoptotic proteins, Bak and Bax, and (2) decreasing the levels of l the anti-apoptotic protein, Bcl-2, compared to cell incubated with a vehicle. Furthermore, DML6, at 5 and 20 μM, induced the cleavage of caspase-9, followed by subsequent cleavage of the executioner caspases, caspase-3 and caspase-7, which produced OV2008 cell death. Overall, our data suggest that DML6 is an apoptosis-inducing compound that should undergo further evaluation as a potential treatment for cervical cancer.

  View details for DOI 10.3390/molecules26144214

  View details for PubMedID 34299490

  View details for PubMedCentralID PMC8306139

• The role of endolysosomal trafficking in anticancer drug resistance. Drug resistance updates : reviews and commentaries in antimicrobial and anticancer chemotherapy Hussein, N. A., Malla, S., Pasternak, M. A., Terrero, D., Brown, N. G., Ashby, C. R., Assaraf, Y. G., Chen, Z. S., Tiwari, A. K. 2021; 57: 100769

  Abstract

  Multidrug resistance (MDR) remains a major obstacle towards curative treatment of cancer. Despite considerable progress in delineating the basis of intrinsic and acquired MDR, the underlying molecular mechanisms remain to be elucidated. Emerging evidences suggest that dysregulation in endolysosomal compartments is involved in mediating MDR through multiple mechanisms, such as alterations in endosomes, lysosomes and autophagosomes, that traffic and biodegrade the molecular cargo through macropinocytosis, autophagy and endocytosis. For example, altered lysosomal pH, in combination with transcription factor EB (TFEB)-mediated lysosomal biogenesis, increases the sequestration of hydrophobic anti-cancer drugs that are weak bases, thereby producing an insufficient and off-target accumulation of anti-cancer drugs in MDR cancer cells. Thus, the use of well-tolerated, alkalinizing compounds that selectively block Vacuolar H⁺-ATPase (V-ATPase) may be an important strategy to overcome MDR in cancer cells and increase chemotherapeutic efficacy. Other mechanisms of endolysosomal-mediated drug resistance include increases in the expression of lysosomal proteases and cathepsins that are involved in mediating carcinogenesis and chemoresistance. Therefore, blocking the trafficking and maturation of lysosomal proteases or direct inhibition of cathepsin activity in the cytosol may represent novel therapeutic modalities to overcome MDR. Furthermore, endolysosomal compartments involved in catabolic pathways, such as macropinocytosis and autophagy, are also shown to be involved in the development of MDR. Here, we review the role of endolysosomal trafficking in MDR development and discuss how targeting endolysosomal pathways could emerge as a new therapeutic strategy to overcome chemoresistance in cancer.

  View details for DOI 10.1016/j.drup.2021.100769

  View details for PubMedID 34217999

• Novel Curcumin-Resveratrol Solid Nanoparticles Synergistically Inhibit Proliferation of Melanoma Cells. Pharmaceutical research Palliyage, G. H., Hussein, N., Mimlitz, M., Weeder, C., Alnasser, M. H., Singh, S., Ekpenyong, A., Tiwari, A. K., Chauhan, H. 2021; 38 (5): 851-871

  Abstract

  Polyphenols such as curcumin (Cur) and resveratrol (Res) have been recently shown to have potential to inhibit proliferation of highly aggressive melanoma cells. This study was designed to investigate the feasibility of a topical delivery system, using a solid lipid nanoparticles (SLNs) loaded delivery systems, that can enhance the skin penetration and anti-cancer efficacy of combination of these polyphenols. Negatively charged Cur-Res SLNs with a mean diameter of 180.2 ± 7.7 nm were prepared using high shear homogenization method. Cur-Res SLNs were found to be stable up to 2 weeks under 4°C. The in vitro release study showed that Res was released five time more than curcumin. The permeability of resveratrol was about 1.67 times that of curcumin from the SLN-gel formulation which was significantly (p < 0.05) lower than from SLN suspension. More than 70% of Cur-Res SLNs were bound to skin locally in a skin binding study suggesting potentially utility of Cur-Res SLNs in the treatment of localized melanoma. In fact, the electrical cell-substrate impedance sensing (ECIS) measurements suggested that Cur-Res combination has potential to stop cell migration of B16F10 melanoma cells. Furthermore, both, Cur-Res SLNs and Cur-Res solution at the ratio of 3:1 demonstrated a strong synergistic inhibition of SK-MEL-28 melanoma cell proliferation. Further evaluation of Cur-Res SLNs in vivo melanoma models are warranted to establish the clinical utility of Cur-Res formulations in melanoma therapy.

  View details for DOI 10.1007/s11095-021-03043-7

  View details for PubMedID 33982225

  View details for PubMedCentralID 6421897

• Endolysosomal trafficking defects leads to caspase independed cell death in colon cancer Hussein, N., Pasternak, M., Kumari, S., Awthe, Z., Gunning, W., Trippier, P. C., Tiwari, A. AMER ASSOC CANCER RESEARCH. 2020

  View details for DOI 10.1158/1538-7445.AM2020-596

  View details for Web of Science ID 000590059303189

• Novel Chrysin-De-Allyl PAC-1 Hybrid Analogues as Anticancer Compounds: Design, Synthesis, and Biological Evaluation. Molecules (Basel, Switzerland) Al-Oudat, B. A., Ramapuram, H., Malla, S., Audat, S. A., Hussein, N., Len, J. M., Kumari, S., Bedi, M. F., Ashby, C. R., Tiwari, A. K. 2020; 25 (13)

  Abstract

  New chrysin-De-allyl-Pac-1 hybrid analogues, tethered with variable heterocyclic systems (4a-4o), were rationally designed and synthesized. The target compounds were screened for in vitro antiproliferative efficacy in the triple-negative breast cancer (TNBC) cell line, MDA-MB-231, and normal human mammary epithelial cells (HMECs). Two compounds, 4g and 4i, had the highest efficacy and selectivity towards MDA-MB-231 cells, and thus, were further evaluated by mechanistic experiments. The results indicated that both compounds 4g and 4i induced apoptosis by (1) inducing cell cycle arrest at the G2 phase in MDA-MB-231 cells, and (2) activating the intrinsic apoptotic pathways in a concentration-dependent manner. Physicochemical characterizations of these compounds suggested that they can be further optimized as potential anticancer compounds for TNBC cells. Overall, our results suggest that 4g and 4i could be suitable leads for developing novel compounds to treat TNBC.

  View details for DOI 10.3390/molecules25133063

  View details for PubMedID 32635530

  View details for PubMedCentralID PMC7412250

• Development of non-apoptotic, caspase-independent cell death inducers against cancer cells Pasternak, M. A., Hussein, N., Chandrabose, K., Erhardt, P. W., Tiwari, A. K. AMER ASSOC CANCER RESEARCH. 2019

  View details for DOI 10.1158/1538-7445.AM2019-2955

  View details for Web of Science ID 000488279401401

• Novel Thienopyrimidine Derivative, RP-010, Induces β-Catenin Fragmentation and Is Efficacious against Prostate Cancer Cells. Cancers Amawi, H., Hussein, N., Boddu, S. H., Karthikeyan, C., Williams, F. E., Ashby, C. R., Raman, D., Trivedi, P., Tiwari, A. K. 2019; 11 (5)

  Abstract

  Thienopyrimidines containing a thiophene ring fused to pyrimidine are reported to have a wide-spectrum of anticancer efficacy in vitro. Here, we report for the first time that thieno[3,2-d]pyrimidine-based compounds, also known as the RP series, have efficacy in prostate cancer cells. The compound RP-010 was efficacious against both PC-3 and DU145 prostate cancer (PC) cells (IC50 < 1 µM). The cytotoxicity of RP-010 was significantly lower in non-PC, CHO, and CRL-1459 cell lines. RP-010 (0.5, 1, 2, and 4 µM) arrested prostate cancer cells in G2 phase of the cell cycle, and induced mitotic catastrophe and apoptosis in both PC cell lines. Mechanistic studies suggested that RP-010 (1 and 2 µM) affected the wingless-type MMTV (Wnt)/β-catenin signaling pathway, in association with β-catenin fragmentation, while also downregulating important proteins in the pathway, including LRP-6, DVL3, and c-Myc. Interestingly, RP-010 (1 and 2 µM) induced nuclear translocation of the negative feedback proteins, Naked 1 and Naked 2, in the Wnt pathway. In addition, RP-010 (0.5, 1, 2 and 4 µM) significantly decreased the migration of PC cells in vitro. Finally, RP-010 did not produce significant toxic effects in zebrafish at concentrations of up to 6 µM. In conclusion, RP-010 may be an efficacious and relatively nontoxic anticancer compound for prostate cancer. Future mechanistic and in vivo efficacy studies are needed to optimize the hit compound RP-010 for lead optimization and clinical use.

  View details for DOI 10.3390/cancers11050711

  View details for PubMedID 31126091

  View details for PubMedCentralID PMC6563099

• Cariprazine, A Dopamine D₂/D₃ Receptor Partial Agonist, Modulates ABCG2-Mediated Multidrug Resistance in Cancer CANCERS Hussein, N., Ashby, C. R., Amawi, H., Nyinawabera, A., Vij, A., Khare, V. M., Karthikeyan, C., Tiwari, A. K. 2018; 10 (9)

  View details for DOI 10.3390/cancers10090308

  View details for Web of Science ID 000448139800032

• Efficient syntheses and anti-cancer activity of xenortides A-D including ent/epi-stereoisomers. Organic & biomolecular chemistry Esmati, N., Maddirala, A. R., Hussein, N., Amawi, H., Tiwari, A. K., Andreana, P. R. 2018; 16 (29): 5332-5342

  Abstract

  A one-pot, two-step, total synthesis of naturally occurring xenortides A, B, C and D, (Xens A-D) isolated from the bacterium Xenorhabdus nematophila, and an entire complementary set of stereoisomers, has been achieved. Compounds were synthesized utilizing an isocyanide-based Ugi 4-CR followed by facile N-Boc deprotection. The reaction sequence took advantage of the chiral pool of N-Boc protected amino acids (l-Leu/Val and d-Leu/Val) with aryl isocyanides, phenyl acetaldehyde and methylamine giving the desired Xens A-D (A and B >98% ee) and all subsequent stereoisomers in reasonable yields upon deprotection followed by separation of diastereomers. Also, detailed mechanistic insights for diastereoselectivity of (-)-Xen A, as a model in the Ugi 4-CR, has been described. Moreover, for the first time, this focused library was screened for cytotoxicity against a panel of epithelial cancer cell lines as well as normal cell lines with an MTT proliferation assay. The structure-activity relationship (SAR) study demonstrated that tryptamides Xen B and D were more active than phenylethylamides Xen A and C. Furthermore, (-)-Xen B (IC50 = 19-25 μM) and ent-(+)-Xen D (IC50 = 21-26 μM) gave the highest cytotoxicity and they were also found to be non-toxic toward normal cells. Importantly, the SAR results indicate that the stereochemistry at C8 and C11 in (-)-Xen B and ent-(+)-Xen D play a critical role in cytotoxic activity.

  View details for DOI 10.1039/c8ob00452h

  View details for PubMedID 29999086

• lH-Pyrazolo[3,4-b]quinolin-3-amine derivatives inhibit growth of colon cancer cells via apoptosis and sub G1 cell cycle arrest. Bioorganic & medicinal chemistry letters Karthikeyan, C., Amawi, H., Viana, A. G., Sanglard, L., Hussein, N., Saddler, M., Ashby, C. R., Moorthy, N. S., Trivedi, P., Tiwari, A. K. 2018; 28 (13): 2244-2249

  Abstract

  A series of lH-pyrazolo[3,4-b]quinolin-3-amine derivatives were synthesized and evaluated for anticancer efficacy in a panel of ten cancer cell lines, including breast (MDAMB-231 and MCF-7), colon (HCT-116, HCT-15, HT-29 and LOVO), prostate (DU-145 and PC3), brain (LN-229), ovarian (A2780), and human embryonic kidney (HEK293) cells, a non-cancerous cell line. Among the eight derivatives screened, compound QTZ05 had the most potent and selective antitumor efficacy in the four colon cancer cell lines, with IC50 values ranging from 2.3 to 10.2 µM. Furthermore, QTZ05 inhibited colony formation in HCT-116 cells in a concentration-dependent manner. Cell cycle analysis data indicated that QTZ05 caused an arrest in the sub G1 cell cycle in HCT-116 cells. QTZ05 induced apoptosis in HCT-116 cells in a concentration-dependent manner that was characterized by chromatin condensation and increase in the fluorescence of fluorochrome-conjugated Annexin V. The findings from our study suggest that QTZ05 may be a valuable prototype for the development of chemotherapeutics targeting apoptotic pathways in colorectal cancer cells.

  View details for DOI 10.1016/j.bmcl.2018.05.045

  View details for PubMedID 29853331

• Bax/Tubulin/Epithelial-Mesenchymal Pathways Determine the Efficacy of Silybin Analog HM015k in Colorectal Cancer Cell Growth and Metastasis. Frontiers in pharmacology Amawi, H., Hussein, N. A., Ashby, C. R., Alnafisah, R., Sanglard, L. M., Manivannan, E., Karthikeyan, C., Trivedi, P., Eisenmann, K. M., Robey, R. W., Tiwari, A. K. 2018; 9: 520

  Abstract

  The inhibition of apoptosis, disruption of cellular microtubule dynamics, and over-activation of the epithelial mesenchymal transition (EMT), are involved in the progression, metastasis, and resistance of colorectal cancer (CRC) to chemotherapy. Therefore, the design of a molecule that can target these pathways could be an effective strategy to reverse CRC progression and metastasis. In this study, twelve novel silybin derivatives, HM015a-HM015k (15a-15k) and compound 17, were screened for cytotoxicity in CRC cell lines. Compounds HM015j and HM015k (15k and 15j) significantly decreased cell proliferation, inhibited colony formation, and produced cell cycle arrest in CRC cells. Furthermore, 15k significantly induced the formation of reactive oxygen species and apoptosis. It induced the cleavage of the intrinsic apoptotic protein (Bax p21) to its more efficacious fragment, p18. Compound 15k also inhibited tubulin expression and disrupted its structure. Compound 15k significantly decreased metastatic LOVO cell migration and invasion. Furthermore, 15k reversed mesenchymal morphology in HCT116 and LOVO cells. Additionally, 15k significantly inhibited the expression of the mesenchymal marker N-cadherin and upregulated the expression of the epithelial marker, E-cadherin. Compound 15k inhibited the expression of key proteins known to induce EMT (i.e., DVL3, β-catenin, c-Myc) and upregulated the anti-metastatic protein, cyclin B1. Overall, in vitro, 15k significantly inhibited CRC progression and metastasis by inhibiting apoptosis, tubulin activity and the EMT pathways. Overall, these data suggest that compound 15k should be tested in vivo in a CRC animal model for further development.

  View details for DOI 10.3389/fphar.2018.00520

  View details for PubMedID 29875662

  View details for PubMedCentralID PMC5974752

• Thienopyrimidine derivatives exert their anticancer efficacy via apoptosis induction, oxidative stress and mitotic catastrophe. European journal of medicinal chemistry Amawi, H., Karthikeyan, C., Pathak, R., Hussein, N., Christman, R., Robey, R., Ashby, C. R., Trivedi, P., Malhotra, A., Tiwari, A. K. 2017; 138: 1053-1065

  Abstract

  In this study, a series of 13 structural variants of thieno[2,3d]pyrimidine derivatives (6a-6m) were synthesized and screened for cytotoxicity in a panel of colorectal, ovarian, and brain cancer cell lines. The selectivity of the compounds was assessed by determining the cytotoxicity in normal epithelial cell line (CHO). The most potent compound, 6j, was efficacious (with IC50 range of 0.6-1.2 μM) in colon (HCT116 and HCT15), brain (LN-229 and GBM-10) and ovarian (A2780 and OV2008) cancer cell lines. In contrast, in the normal cell line (CHO), the IC50 values for 6j were 14 ± 1.3 μM. Compound 6j significantly inhibited the clonogenic potential of HCT116, OV2008 and A2780 cell lines in concentration - dependent (0.5-4 μM) manner. Also, 6j induced 1) formation of reactive oxygen species; 2) apoptosis and 3) mitotic catastrophe in HCT116 and OV2008 cells (IC50 = 0.5-2 μM). Furthermore, apoptosis was the predominant mechanism of death in A2780 cells. The cytotoxicity of 6j in wild type HCT116 cells was similar to that in HCT116 cells lacking the apoptotic genes for Bax, Bak, or Bak and Bax, indicating that 6j induces mitotic catastrophe as alternative mechanism of death when when certain apoptotic proteins are absent. In summary, this study has identified a lead molecule, 6j, that selectively induces oxidative stress, apoptosis and mitotic catastrophe in specific cancer (colon and ovarian) cell lines.

  View details for DOI 10.1016/j.ejmech.2017.07.028

  View details for PubMedID 28759878

• HM015k, a Novel Silybin Derivative, Multi-Targets Metastatic Ovarian Cancer Cells and Is Safe in Zebrafish Toxicity Studies. Frontiers in pharmacology Amawi, H., Hussein, N. A., Karthikeyan, C., Manivannan, E., Wisner, A., Williams, F. E., Samuel, T., Trivedi, P., Ashby, C. R., Tiwari, A. K. 2017; 8: 498

  Abstract

  This study was designed to determine the in vitro mechanisms by which the novel silybin derivative, (E)-3-(3-(benzyloxy) phenyl)-1-(4-hydroxyphenyl)prop-2-en-1-one (HM015k or 15k), produces its anticancer efficacy in ovarian cancer cells. Compound 15k induced apoptosis in ovarian cancer cells in a time-dependent manner by significantly upregulating the expression of Bax and Bak and downregulating the expression of Bcl-2. Interestingly, 15k induced the cleavage of Bax p21 into its more efficacious cleaved form, Bax p18. In addition, caspase 3 and caspase 9 were cleaved to their active forms, inducing the cleavage of poly ADP ribose polymerase (PARP) and β-catenin. Furthermore, in OV2008 cells, 15k induced significant cleavage in nuclear β-catenin to primarily inactive fragments of lower molecular weight. Furthermore, 15k reversed the metastatic potential of OV2008 cells by inhibiting their migration and invasiveness. The mesenchymal phenotype in OV2008 was reversed by 15k, causing cells to be rounder with epithelial-like phenotypes. The 15k-induced reversal was further confirmed by significant upregulation of the E-cadherin expression, an epithelial marker, while N-cadherin, a mesenchymal marker, was downregulated in OV2008 cells. Compound 15k inhibited the expression of the oncogenic c-Myc protein, downregulated proteins DVL3 and DVL2 and significantly upregulated cyclin B1. Also, 15k significantly downregulated the expression levels of ABCG2 and ABCB1 transporters in resistant ABCG2 overexpressing H460/MX20 and resistant ABCB1 overexpressing MDCK/MDR1 cells, respectively. Finally, 15k was safe in zebrafish in vivo model at concentrations up to 10 μM and induced no major toxicities in cardiac, morphology and swimming position parameters. Overall, 15k is a multi-targeted inhibitor with efficacy against metastatic and resistant ovarian cancer. Future in vivo studies will be conducted to determine the efficacy of 15k in tumor-bearing animals.

  View details for DOI 10.3389/fphar.2017.00498

  View details for PubMedID 28824426

  View details for PubMedCentralID PMC5539246

• Novel sylibin analogues target ovarian cancer EMT-Wnt/β-catenin resistance pathways Amawi, H. A., Hussein, N., Fetcenkoa, A., Alnafisah, R., Chandrabose, K., Manivannan, E., Trivedi, P., Tiwari, A. K. AMER ASSOC CANCER RESEARCH. 2017

  View details for DOI 10.1158/1538-7445.AM2017-2238

  View details for Web of Science ID 000442496705094

• The dopamine (DA) D₃ receptor antagonists (PG01037, NGB2904, SB-277011A, and U99194) significantly attenuate ABCG2-mediated multidrug resistance Hussein, N., Amawi, H., Ashby, C. R., Chandrabose, K., Mittal, R., Christman, R., Trivedi, P., Tiwari, A. AMER ASSOC CANCER RESEARCH. 2017

  View details for DOI 10.1158/1538-7445.AM2017-1191

  View details for Web of Science ID 000442496703114

• The dopamine D3 receptor antagonists PG01037, NGB2904, SB277011A, and U99194 reverse ABCG2 transporter-mediated drug resistance in cancer cell lines. Cancer letters Hussein, N., Amawi, H., Karthikeyan, C., Hall, F. S., Mittal, R., Trivedi, P., Ashby, C. R., Tiwari, A. K. 2017; 396: 167-180

  Abstract

  The ATP - binding cassette (ABC) family G2 (ABCG2) transporters are known to produce multidrug resistance (MDR) in cancer, thereby limiting the clinical response to chemotherapy. Molecular modeling data indicated that certain dopamine (DA) D3 receptor antagonists had a significant binding affinity for ABCG2 transporter. Therefore, in this in vitro study, we determined the effect of the D3 receptor antagonists PG01037, NGB2904, SB277011A, and U99194 on MDR resulting from the overexpression of ABCG2 transporters. The D3 receptor antagonists, at concentrations >100 μM, did not significantly affect the viability of H460-MX20, S1M1-80, A549-MX10 or wild type ABCG2 overexpressing (HEK293-R2) cells. However, at concentrations ranging from 0.01 to 10 μM, the D3 receptor antagonists PG01037, NGB2904, SB-277011A, and U99194 significantly increased the efficacy of the anticancer drugs mitoxantrone and doxorubicin in ABCG2-overexpressing MDR cells. Efflux studies indicated that both PG01037 and NGB2904, at a concentration of 5 μM, significantly decreased the efflux of rhodamine 123 from H460-MX20 cells. Interestingly, 5 μM of PG01037 or NGB2904 significantly decreased the expression levels of the ABCG2 protein, suggesting that these compounds inhibit both the function and expression of ABCG2 transporters at non-toxic concentrations.

  View details for DOI 10.1016/j.canlet.2017.03.015

  View details for PubMedID 28323029

• Design and discovery of silybin analogues as antiproliferative compounds using a ring disjunctive - Based, natural product lead optimization approach. European journal of medicinal chemistry Manivannan, E., Amawi, H., Hussein, N., Karthikeyan, C., Fetcenko, A., Narayana Moorthy, N. S., Trivedi, P., Tiwari, A. K. 2017; 133: 365-378

  Abstract

  The present study reports the synthesis and anticancer activity evaluation of twelve novel silybin analogues designed using a ring disjunctive-based natural product lead (RDNPL) optimization approach. All twelve compounds were tested against a panel of cancer cells (i.e. breast, prostate, pancreatic, and ovarian) and compared with normal cells. While all of the compounds had significantly greater efficacy than silybin, derivative 15k was found to be highly potent (IC50 < 1 μM) and selective against ovarian cancer cell lines, as well as other cancer cell lines, compared to normal cells. Preliminary mechanistic studies indicated that the antiproliferative efficacy of 15k was mediated by its induction of apoptosis, loss of mitochondrial membrane potential and cell cycle arrest at the sub-G1 phase. Furthermore, 15k inhibited cellular microtubules dynamic and assembly by binding to tubulin and inhibiting its expression and function. Overall, the results of the study establish 15k as a novel tubulin inhibitor with significant activity against ovarian cancer cells.

  View details for DOI 10.1016/j.ejmech.2017.03.033

  View details for PubMedID 28411546

• A reappraisal of the 6-O-desmethylnaproxen-sulfating activity of the human cytosolic sulfotransferases. Canadian journal of physiology and pharmacology Alherz, F. A., Almarghalani, D. A., Hussein, N. A., Kurogi, K., Liu, M. C. 2017; 95 (6): 647-651

  Abstract

  In this study, we aimed to obtain a comprehensive account of the human cytosolic sulfotransferases (SULTs) that are capable of sulfating 6-O-desmethylnaproxen (O-DMN), a major metabolite of naproxen. Of the 13 known human SULTs tested, 7 (SULT1A1, SULT1A2, SULT1A3, SULT1B1, SULT1C2, SULT1C4, and SULT1E1) displayed O-DMN-sulfating activity, when analyzed using an elevated substrate concentration (500 μmol·L-1) together with 14 μmol·L-1 of the sulfate donor, 3'-phosphoadenosine-5'-phosphosulfate (PAPS). At 10 μmol·L-1 O-DMN concentration, however, only SULT1A1 and SULT1A3 displayed detectable activity, with the former being nearly 2 orders of magnitude more active than the latter. A pH-dependence study indicated that SULT1A1 exhibited a broad pH optimum spanning pH 5.5-7. Kinetic parameters of the sulfation of O-DMN by SULT1A1 were determined. The production and release of sulfated O-DMN was demonstrated using cultured human HepG2 hepatoma cells and Caco-2 colon carcinoma cells. Moreover, assays using human organ specimens revealed that the O-DMN-sulfating activities present in the cytosols of liver and small intestine (at 502.5 and 497.2 pmol·min-1·(mg protein)-1, respectively) were much higher than those detected for the cytosols of lung and kidney. Taken together, these results provided relevant information concerning the sulfation of O-DMN both in vitro and in vivo.

  View details for DOI 10.1139/cjpp-2016-0403

  View details for PubMedID 28177672