Biology subjects

Civanelli, E.

Publications and source records attributed to Civanelli, E..

3 recordsLinked to original sources

bioRxiv2025

Dual targeting of BTK and BCL2 enhances apoptosis in marginal zone lymphoma models: preclinical activity of BGB-16673 and sonrotoclax

BackgroundBTK and BCL2 represent key therapeutic targets in B-cell lymphomas, including marginal zone lymphoma (MZL). Here, we evaluated the novel BTK degrader BGB-16673, and the second-generation BCL2 inhibitor sonrotoclax in a panel of MZL cell lines, including models with acquired resistance to BTK, BCL2 and PI3K inhibitors, as single agents and in combination. MethodsCytotoxicity, transcriptomic changes, apoptosis, and protein expression were assessed for BGB-16673 alone and in combination with venetoclax, sonrotoclax, bendamustine, selinexor, lenalidomide, and tazemetostat. ResultsBGB-16673 demonstrated single-agent activity in selected MZL cell lines, inducing BTK degradation and transcriptional repression of BCR signaling and MYC targets. Its transcriptional and phenotypic effects overlapped with those of zanubrutinib but also included specific modulation of oxidative phosphorylation genes. Combination studies revealed that BGB-16673 synergized with other targeted agents, achieving the best results with sonrotoclax, venetoclax, bendamustine, lenalidomide, and rituximab. The mechanism of action of the combination with the BCL2 inhibitors was further evaluated. The combination with sonrotoclax consistently enhanced apoptosis across MZL models, outperforming venetoclax in potency in most cell lines. Sonrotoclax showed over 10-fold higher activity than venetoclax in three MZL lines and demonstrated synergistic interaction with BGB-16673. Mechanistically, the combination leads to BTK degradation and modulation of anti-apoptotic proteins such as BCL2, MCL1, and BCL-XL. ConclusionOur findings underscore the potential of BGB-16673 as a therapeutic agent, both as a monotherapy and in combination regimens, for treating MZL patients. Additionally, the results also identify the second-generation BCL2 inhibitor sonrotoclax as another drug to be explored in the same patient populations.

cancer biology↗

bioRxiv2025

Characterization of lymphoma models for the surface ROR1 expression

Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is a developmental antigen aberrantly expressed in several B-cell malignancies and represents an attractive therapeutic target. Here, we systematically characterized ROR1 expression in a large panel of B-cell lymphoma cell lines and assessed the activity of a ROR1-targeting antibody-drug conjugate (ADC). ROR1 RNA expression was first evaluated using a previously generated total RNA-Seq dataset from 47 B-cell lymphoma cell lines. ROR1 transcripts were detectable in most models, with 19 cell lines showing moderate to high expression. To determine whether ROR1 is present on the cell surface, we next analyzed 29 cell lines by flow cytometry using a PE-conjugated anti-ROR1 antibody. Only a subset of cell lines expressed appreciable levels of ROR1 on the cell membrane, with the highest expression observed in mantle cell lymphoma (MCL) models compared with other B-cell lymphoma subtypes, and detectable expression in a few diffuse large B-cell lymphoma (DLBCL) and one marginal zone lymphoma (MZL) line. ROR1 RNA and surface protein levels were highly correlated. As a proof of principle, four ROR1-positive DLBCL cell lines and one cell line with low surface protein were exposed to the ROR1-targeting ADC zilovertamab vedotin for 120 hours. All ROR1-positive models were sensitive, with IC50 values ranging from 28 to 58 nM, whereas the remaining one was largely resistant. In summary, we defined a set of well-characterized lymphoma models suitable for preclinical studies of ROR1-directed agents, confirming the functional relevance of ROR1 surface expression for the activity of zilovertamab vedotin. The gene coding for the receptor tyrosine kinase-like orphan receptor 1 (ROR1) was initially identified while looking for genes encoding proteins with tyrosine kinase-like domains 1,2. It is now understood that ROR1 possesses intrinsic tyrosine kinase activity and also functions as a receptor for Wnt5a, which activates {beta}-catenin-independent non-canonical pathways, sustaining cell survival and oncogenesis 2,3. In adult tissues, including normal B lymphocytes, ROR1 is preferentially expressed by cancer cells rather than normal cells and has a role in sustaining the growth and survival of neoplastic cells 2,4. Several therapeutic strategies targeting ROR1 have been developed, including naked monoclonal antibodies, antibody-drug conjugates (ADCs), small molecule inhibitors, bispecific T cell engager (BiTE), and chimeric antigen receptor (CAR) T cells have been developed 2, with some currently in advanced clinical evaluation 2. Here, we characterized a vast panel of lymphoma cell lines for their ROR1 RNA level and ROR1 cell surface protein expression, and showed, using zilovertamab vedotin as proof-of-principle, that these models can be exploited to study this promising class of anti-cancer agents.

cancer biology↗

bioRxiv2025

The anti-metabolite KAT/3BP has in vitro and in vivo anti-tumor activity in lymphoma models

Reprogramming of cellular metabolism is a hallmark of cancer, offering therapeutic opportunities to target cancer cell vulnerabilities for therapeutic purposes. 3-Bromopyruvate (3BP), a small alkylating agent, acts as an anti-metabolite to vital substrates in cancer metabolism and exhibits antitumor activity across various cancer types, but the unformulated 3BP can cause high toxicity. This study explores the efficacy of the 3BP clinical derivative KAT/3BP, currently in phase 1 for patients with hepatocellular carcinoma, in lymphoma models. In vitro, KAT/3BP demonstrated cytotoxicity across 12 lymphoma cell lines, including diffuse large B-cell lymphoma and mantle cell lymphoma, with a median IC50 of 3.7 M. KAT/3BP was also effective against lymphoma cell lines with acquired resistance to FDA-approved therapies. In vivo, we observed reduced tumor size in a syngeneic mouse model, with the combination of oral and intratumoral administration proving most effective. Additionally, KAT/3BP exhibited synergistic activity when combined with lymphoma therapies, including bendamustine and R-CHOP. These findings underscore the potential of KAT/3BP as a novel therapeutic option for a single agent or a combination of lymphomas.

cancer biology↗