Andrius Kleinauskas

• Stenberg, Vilde Yuli; Liukaityte, Rugile; Kristiansen, Sandra Kvarsvik; Wangen-Riise, Rina; Kleinauskas, Andrius & Dunne, Victoria [Vis alle 7 forfattere av denne artikkelen] (2025). Cytotoxicity and cell cycle changes in prostate cancer cells with differing PSMA expression and p53 status after treatment with PSMA-targeting radioligand [212Pb]Pb-AB001. Scientific Reports. 16(1). doi: 10.1038/s41598-025-29785-7. Fulltekst i vitenarkiv Vis sammendrag

  Targeted alpha therapy holds promise for treating advanced prostate cancer, but the interplay between prostate-specific membrane antigen (PSMA) expression, p53 status, and downstream cell fate remains poorly defined. This study evaluates the cytotoxic and cell cycle effects of the alpha-emitting radioligand [212Pb]Pb-AB001 in prostate cancer cell lines with differing PSMA expression and p53 status: C4-2 (PSMA+/TP53-wild-type) and PC-3 PIP (PSMA+++/ TP53-null). [212Pb]Pb-AB001 significantly inhibited proliferation and clonogenic survival in both cell lines in an activity-dependent manner. At 95% clonogenic inhibition, both cell lines exhibited G2-phase arrest, S-phase suppression and reduced mitotic entry on day 1. At higher activities, PC-3 PIP cells showed polyploidy, and features consistent with mitotic catastrophe and senescence. Cytotoxicity was more pronounced in C4-2 3D spheroid models than in 2D monolayers, suggesting contribution of crossfire and bystander effects. Total cell-bound activity, rather than added activity, better predicted radiotoxicity in both TP53-wild-type and TP53-null cell lines, indicating that its therapeutic effect is primarily governed by PSMA-mediated uptake rather than p53 status. These results support the therapeutic potential of [212Pb]Pb-AB001 across cells with varying TP53 status and suggest that combining [212Pb]Pb-AB001 with DNA repair or checkpoint inhibitors may enhance treatment efficacy.

• Liukaityte, Rugile; Stenberg, Vilde Yuli; Kleinauskas, Andrius; Juzenas, Petras; Urbanucci, Alfonso & Juzeniene, Asta (2025). Combination of PSMA targeting alpha-emitting radioligand [212Pb]Pb-AB001 with BET bromodomain inhibitors in in vitro prostate cancer models. Medical Oncology (MO). ISSN 1357-0560. 42(8). doi: 10.1007/s12032-025-02925-9. Fulltekst i vitenarkiv Vis sammendrag

  The alpha-emitting radioligand [ 212 Pb]Pb-AB001, targeting prostate-specific membrane antigen (PSMA), is a promising therapy approach for prostate cancer. Bromodomain and extra-terminal (BET) protein inhibitors, such as AZD5153 and JQ1, disrupt oncogenic transcriptional programs by altering chromatin structure. This study evaluated whether BET inhibition enhances the efficacy of radioligand therapy. Cytotoxic effects of [ 212 Pb]Pb-AB001 alone and in combination with BET inhibitors were assessed in 2D monolayers and a 3D spheroid model of PSMA-positive C4-2 prostate cancer cells. Cell viability, cell cycle alterations, and DNA damage were assessed using viability assays and flow cytometry. Spheroid growth and viability were assessed by fluorescence microscopy. AZD5153 was more potent than JQ1 in reducing cell proliferation. [ 212 Pb]Pb-AB001 induced activity- and time-dependent cytotoxicity with a delayed apoptotic response. BET inhibitors induced G1 arrest, while [ 212 Pb]Pb-AB001 caused G2 arrest. Combination treatment reduced cell viability in an additive manner but did not further affect cell cycle distribution or increase apoptosis compared with [ 212 Pb]Pb-AB001 alone. γH2AX staining in 2D models showed an activity- and time-dependent increase in DNA damage at 1, 3 and 6 days post-treatment with [ 212 Pb]Pb-AB001. BET inhibitors alone induced minimal γH2AX, and combination treatments did not enhance DNA damage beyond [ 212 Pb]Pb-AB001 alone. In 3D spheroids, combination treatment led to synergistic growth suppression. In conclusion, these findings indicate that therapeutic inhibition of BET bromodomain in combination with the alpha-emitting radioligand [ 212 Pb]Pb-AB001 could significantly enhance tumour control and should be further evaluated in metastatic prostate cancer models.

• Espeland, Kristian; Christensen, Eidi; Aandahl, Astrid; Ulvær, Andreas; Warloe, Trond & Kleinauskas, Andrius [Vis alle 11 forfattere av denne artikkelen] (2024). Extracorporeal Photopheresis with 5-Aminolevulinic Acid in Crohn’s Disease—A First-in-Human Phase I/II Study. Journal of Clinical Medicine. 13:6198(20), s. 1–12. doi: 10.3390/jcm13206198. Fulltekst i vitenarkiv Vis sammendrag

  Background/Objectives: With the increasing prevalence of Crohn’s disease (CD), treatment options for patients who fail conventional and advanced therapy are highly needed. Therefore, we explored the safety and efficacy of extracorporeal photopheresis (ECP) using 5-aminolevulinic acid (ALA) and blue light (405 nm). Methods: Patients with active CD who failed or were intolerant to biological therapy were eligible. Mononuclear cells (90 mL) were collected from each patient using a Spectra Optia® apheresis system and diluted with 100 mL of 0.9% sodium chloride in a collection bag. The cells were incubated with ALA at a concentration of 3 millimolar (mM) for 60 min ex vivo and illumination with an LED blue light (405 nm) source (BLUE-PIT®) before reinfusion to the patient. Recording of vital signs and adverse events were regularly performed. At week 13, we assessed the patients with colonoscopy, the Harvey Bradshaw Index (HBI), the Inflammatory Bowel disease Health Related Quality of Life Questionnaire, and the measurement of serum C-reactive protein and fecal calprotectin (FC) levels. Biopsies of the intestines were taken for immunohistochemistry. Results: Seven patients were included. Four patients completed the treatments, with a total of 24 treatments. Three of the four patients achieved a favorable response, including a lower HBI, lower FC levels, and/or endoscopic improvement. No significant adverse events were observed. The remaining three patients received only one, three, or five treatments due to technical difficulties, medical reasons, or the withdrawal of informed consent. Conclusions: ALA-based ECP appears safe and seems to give some clinical improvement for the patients with active CD who failed to respond to conventional and advanced therapies.

• Espeland, Kristian; Kleinauskas, Andrius; Juzenas, Petras; Darvekar, Sagar Ramesh; Vasovic, Vlada & Warloe, Trond [Vis alle 9 forfattere av denne artikkelen] (2023). Photodynamic effects with 5-aminolevulinic acid on cytokines and exosomes in human peripheral blood mononuclear cells from patients with Crohn’s disease. International Journal of Molecular Sciences. ISSN 1661-6596. 24(5), s. 1–20. doi: 10.3390/ijms24054554. Fulltekst i vitenarkiv Vis sammendrag

  Photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA) which is the precursor of the photosensitizer protoporphyrin IX (PpIX) is an available treatment for several diseases. ALA-PDT induces the apoptosis and necrosis of target lesions. We have recently reported the effects of ALA-PDT on cytokines and exosomes of human healthy peripheral blood mononuclear cells (PBMCs). This study has investigated the ALA-PDT-mediated effects on PBMC subsets from patients with active Crohn’s disease (CD). No effects on lymphocyte survival after ALA-PDT were observed, although the survival of CD3−/CD19+ B-cells seemed slightly reduced in some samples. Interestingly, ALA-PDT clearly killed monocytes. The subcellular levels of cytokines and exosomes associated with inflammation were widely downregulated, which is consistent with our previous findings in PBMCs from healthy human subjects. These results suggest that ALA-PDT may be a potential treatment candidate for CD and other immune-mediated diseases.

• Espeland, Kristian; Kleinauskas, Andrius; Juzenas, Petras; Brech, Andreas; Darvekar, Sagar & Vasovic, Vlada [Vis alle 10 forfattere av denne artikkelen] (2022). Photodynamic Effects with 5-Aminolevulinic Acid on Cytokines and Exosomes in Human Peripheral Blood Mononuclear Cells. Biomedicines. 10(2). doi: 10.3390/biomedicines10020232. Fulltekst i vitenarkiv Vis sammendrag

  Photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA), a precursor to the potent photosensitizer, protoporphyrin IX (PpIX), is an established modality for several malignant and premalignant diseases. This treatment is based on the light-activated PpIX in targeted lesions. Although numerous studies have confirmed the necrosis and apoptosis involved in the mechanism of action of this modality, little information is available for the change of exosome levels after treatment. We report from the first study on the effects of ALA-PDT on cytokines and exosomes of human healthy peripheral blood mononuclear cells (PBMCs). The treatment reduced the cytokines and exosomes studied, although there was variation among individual PBMC samples. This reduction is consistent with PDT-mediated survivals of subsets of PBMCs. More specifically, the ALA-PDT treatment apparently decreased all pro-inflammatory cytokines included, suggesting that this treatment may provide a strong anti-inflammatory effect. In addition, the treatment has decreased the levels of different types of exosomes, the HLA-DRDPDQ exosome in particular, which plays an important role in the rejection of organ transplantation as well as autoimmune diseases. These results may suggest future therapeutic strategies of ALA-PDT.

• Sioud, Mouldy; Juzenas, Petras; Zhang, Qindong; Kleinauskas, Andrius & Qian, Peng (2021). Evaluation of in vitro phototoxicity of a minibody-ir700 conjugate using cell monolayer and multicellular tumor spheroid models. Cancers. 13(13). doi: 10.3390/cancers13133356. Fulltekst i vitenarkiv
• Darvekar, Sagar; Juzenas, Petras; Oksvold, Morten Pedersen; Kleinauskas, Andrius; Holien, Toril & Christensen, Eidi [Vis alle 9 forfattere av denne artikkelen] (2020). Selective Killing of Activated T Cells by 5-Aminolevulinic Acid Mediated Photodynamic Effect: Potential Improvement of Extracorporeal Photopheresis. Cancers. 12. doi: 10.3390/cancers12020377. Fulltekst i vitenarkiv Vis sammendrag

  Extracorporeal photopheresis (ECP), a modality that exposes isolated leukocytes to the photosensitizer 8-methoxypsoralen (8-MOP) and ultraviolet-A (UV-A) light, is used to treat conditions such as cutaneous T-cell lymphoma and graft-versus-host disease. However, the current procedure of ECP has limited selectivity and efficiency; and produces only partial response in the majority of treated patients. Additionally, the treatment is expensive and time-consuming, so the improvement for this modality is needed. In this study, we used the concept of photodynamic therapy (PDT) with 5-aminolevulinic acid (ALA), a precursor of an endogenously synthesized photosensitizer protoporphyrin IX (PpIX) in combination with blue light to explore the possibility of targeting activated human blood T cells ex vivo. With various T-cell activation protocols, a high ALA-induced PpIX production took place in activated CD3+, CD4+CD25+, and CD8+ T cell populations with their subsequent killing after blue light exposure. By contrast, resting T cells were much less damaged by the treatment. The selective and effective killing effect on the activated cells was also seen after co-cultivating activated and resting T cells. Under our clinically relevant experimental conditions, ALA-PDT killed activated T cells more selectively and efficiently than 8-MOP/UV-A. Monocyte-derived dendritic cells (DCs) were not affected by the treatment. Incubation of ALA-PDT damaged T cells with autologous DCs induced a downregulation of the co-stimulatory molecules CD80/CD86 and also upregulation of interleukin 10 (IL-10) and indoleamine 2,3-dioxygenase expression, two immunosuppressive factors that may account for the generation of tolerogenic DCs. Overall, the data support the potential use of ALA-PDT strategy for improving ECP by selective and effective killing of activated T cells and induction of immune tolerance.

• Juzenas, Petras; Kleinauskas, Andrius; Luo, Pengju George & Sun, Ya-Ping (2013). Photoactivatable carbon nanodots for cancer therapy. Applied Physics Letters. ISSN 0003-6951. 103(6). doi: 10.1063/1.4817787. Fulltekst i vitenarkiv
• Kleinauskas, Andrius; Rocha, Sandra; Sahu, Sushant; Sun, Ya-Ping & Juzenas, Petras (2013). Carbon-core silver-shell nanodots as sensitizers for phototherapy and radiotherapy. Nanotechnology. ISSN 0957-4484. 24(32). doi: 10.1088/0957-4484/24/32/325103. Fulltekst i vitenarkiv

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