Anna Maria Nowicka

Integrins are transmembrane receptors that mediate bidirectional signaling across the plasma membrane and play a crucial role in tumor progression, metastasis, and cellular communication. In this study, we performed a comparative structural and biophysical analysis of the PTHTRWA-functionalized extracellular vesicles (PTHTRWA-EVs) interacting with α 2 β 1 and α 5 β 1 integrins to investigate the molecular determinants underlying selective recognition. Surface plasmon resonance experiments were used to characterize multivalent bioengineered EVs --- integrin binding under physiological conditions, while molecular dynamics simulations provided residue-level insight into local ligand-receptor interaction patterns and conformational preferences. These results indicate that PTHTRWA binding is associated with local conformational rearrangements consistent with stabilization of an open-like binding geometry at the integrin interface. Together, these complementary approaches highlight the potential of PTHTRWA-functionalized EVs as a platform for targeted drug delivery and cancer diagnostics.

Magnetic fluid hyperthermia (MFH) emerges as a potential new therapeutic strategy for the treatment of lung cancer. However, the biological endpoints underpinning its therapeutic efficacy remain insufficiently defined in this tumor. In this work, we advance the translational potential of MFH by delineating metabolic, structural and biophysical endpoints in patient-derived lung cancer models. Specifically, we evaluated the effects of MFH using Mg_0.1-γ-Fe₂O₃(mPEG-silane)_0.5 nanoparticles (NPs) subjected to an alternating magnetic field (AMF) on patient-derived lung cancer cells (in vitro) and NUDE Balb/c mice bearing patient-derived lung cancer xenografts, PDX (in vivo). We elucidated that MFH induces metabolic dysfunction in lung cancer cells, leading to reduced proliferation, diminished colony formation and restricted cell migration. Moreover, alterations in metallomic profiles and changes in glycan structures were detected in lung cancer cells treated with MFH. These were accompanied by released matrix metalloproteinases (MMP-1, MMP-2, and MMP-9) and increased cell membrane permeability, indicating that the primary effect of MFH on human lung cancer cells targets membrane integrity and the cell–extracellular matrix environment. Studies have shown that mice bearing lung cancer PDX subjected to MFH experienced a significant reduction in tumor growth compared to the untreated control. The CEM43 value, representing the cumulative equivalent minutes at 43 °C, was estimated to be approximately 9.1 minutes in MFH-treated animals, while the specific absorption rate (SAR) ranged between 389 and 475 W g⁻¹. Together, these findings refine the characterization of hyperthermia treatment endpoints and demonstrate magnetic fluid hyperthermia as a promising translational approach for lung cancer therapy, meriting further evaluation in future clinical applications.

Studies of (S)-thalidomide were conducted on zebrafish embryos subjected to electroporation processes using a square wave pulse generator. The results showed that the electroporation increases the absorption of (S)-thalidomide through the chorion into the growing embryos, which was confirmed by increased thalidomide levels in the embryo bodies at different hours post-treatments using differential pulse voltammetry and controlled-growth mercury drop electrode techniques. (S)-thalidomide administered by electroporation produced structural body deformations in zebrafish embryos as showed by scanning electron microscopy studies. Detailed transmission electron microscopy analysis evidenced multiple deposits of the outer layer and translucencies in the chorion structure, which was also pronounced on the mitochondrial membranes. The results confirmed the spontaneous conversion of the (S)-thalidomide enantiomer to the (R)-enantiomer in embryos exposed to the (S)-thalidomide only and subjected to electroporation pulses. The electroporation was found as a promising method to increase the uptake of (S)-thalidomide in the developmental studies on early zebrafish embryos growing in the chorion.

The 1,3-dipolar cycloaddition reaction (click chemistry approach) was employed to create a hexa-ferrocenylated 1,3,5-triphenylbenzene derivative. Leveraging the presence of metal-chelating sites associated with 1,2,3-triazole moieties and 1,4-dinitrogen systems (ethylenediamine-like), as well as tridentate chelating sites (1,4,7-trinitrogen, diethylene triamine-like) systems, the application of this molecule as a chemosensor for divalent transition metal cations was investigated. The interactions were probed voltammetrically and spectrofluorimetrically against seven selected cations: iron(II) (Fe²⁺), cobalt(II) (Co²⁺), nickel(II) (Ni²⁺), copper(II) (Cu²⁺), zinc(II) (Zn²⁺), cadmium(II) (Cd²⁺), and manganese(II) (Mn²⁺). Electrochemical assays revealed good detection properties, with very low limits of detection (LOD), for Co²⁺, Cu²⁺, and Cd²⁺ in aqueous solution (0.03-0.09 μM). Emission spectroscopy experiments demonstrated that the title compound exhibited versatile detection properties in solution, specifically turn-off fluorescence behavior upon the addition of each tested transition metal cation. The systems were characterized by satisfactory Stern-Volmer constant values (10⁵-10⁶ M^-1) and low LOD, especially for Zn²⁺ and Co²⁺ (at the nanomolar concentration level).

Background: Traditional small-molecule chemotherapeutics usually do not distinguish tumors from healthy tissues. However, nanotechnology creates nanocarriers that selectively deliver drugs to their site of action. This work is the next step in the development of the quantum dot−β−cyclodextrin−folic acid (QD−β−CD−FA) platform for targeted and selected delivery of C−2028 unsymmetrical bisacridine in cancer therapy. Methods: Herein, we report an initial biological evaluation (using flow cytometry and light microscopy) as well as cell migration analysis of QD−β−CD(C−2028)−FA nanoconjugate and its components in the selected human lung and prostate cancer cells, as well as against their respective normal cells. Results: C−2028 compound induced apoptosis, which was much stronger in cancer cells compared to normal cells. Conjugation of C−2028 with QD_green increased cellular senescence, while the introduction of FA to the conjugate significantly decreased this process. C−2028 nanoencapsulation also reduced cell migration. Importantly, QD_green and QD_green−β−CD−FA themselves did not induce any toxic responses in studied cells. Conclusions: In conclusion, the results demonstrate the high potential of a novel folic acid-targeted receptor quantum dot−β−cyclodextrin carrier (QD_green−β−CD−FA) for drug delivery in cancer treatment. Nanoplatforms increased the amount of delivered compounds and demonstrated high suitability.

Personalized medicine is a new approach to modern oncology. Here, to facilitate the application of extracellular vesicles (EVs) derived from lung cancer cells as potent advanced therapy medicinal products in lung cancer, the EV membrane was functionalized with a specific ligand for targeting purposes. In this role, the most effective heptapeptide in binding to lung cancer cells (PTHTRWA) was used. The functionalization process of EV surface was performed through the C- or N-terminal end of the heptapeptide. To prove the activity of the EVs functionalized with PTHTRWA, both a model of lipid membrane mimicking normal and cancerous cell membranes as well as human adenocarcinomic alveolar basal epithelial cells (A549) and human normal bronchial epithelial cells (BEAS-2B) have been exposed to these bioconstructs. Magnetic resonance imaging (MRI) showed that the as-bioengineered PTHTRWA-EVs loaded with superparamagnetic iron oxide nanoparticle (SPIO) cargos reach the growing tumor when dosed intravenously in NUDE Balb/c mice bearing A549 cancer. Molecular dynamics (MD) in silico studies elucidated a high affinity of the synthesized peptide to the α5β1 integrin. Preclinical safety assays did not evidence any cytotoxic or genotoxic effects of the PTHTRWA-bioengineered EVs.

Atenolol β‐blocker drug (ATN) to treat hypertension and cardiovascular disorders such as angina pectoris is manufactured commercially in a racemic form, while only its (S)‐enantiomer shows selective β1‐blocking activity. Here, a new axially chiral thiolated naphthalene diimide derivatives (HS‐BIN‐NDI) for the rapid, simple, and sensitive detection of ATN enantiomers is presented. The Surface Plasmon Resonance (SPR) and Atomic Force Microscopy (AFM) experiments show that interactions between a chiral selector and a chiral analyte occur only when their configurations are opposite. The extensive computational simulations support this finding. The association coefficient determined by SPR is high (10⁵‐10⁶ M⁻¹ s⁻¹) and indicates that the interactions are electrostatic. The analytical utility of atropisomeric HS‐BIN‐NDIs is proven, and the voltammetric sensors designed to determine ATN enantiomers are constructed. The developed sensors are characterized by a wide analytical operating range from 5.0·10⁻³ to 1.0 µm, nanomolar detection limit, high selectivity, and a long lifetime. Several additives like urea, creatinine, glucose, human albumin, and hemoglobin do not affect ATN sensing.