https://hdl.handle.net/10481/32741 2026-04-05T04:42:13Z 2026-04-05T04:42:13Z Pérez, Juana M Rodríguez Criado, Jorge Quiñonero Muñoz, Francisco José Gutiérrez Segura, Natalia López-Vargas, Mireya E. Forte-Castro, Arantxa Ruiz Muelle, Ana Belén Salcedo-Abraira, Pablo Rodríguez Diéguez, Antonio Melguizo Alonso, Consolación Prados Salazar, José Carlos Fernández, Ignacio https://hdl.handle.net/10481/110911 2026-02-12T09:31:06Z

Chiral amino acid–based anthraquinones: synthesis, structural characterization, and structure–activity relationships in cancer cell lines Pérez, Juana M; Rodríguez Criado, Jorge; Quiñonero Muñoz, Francisco José; Gutiérrez Segura, Natalia; López-Vargas, Mireya E.; Forte-Castro, Arantxa; Ruiz Muelle, Ana Belén; Salcedo-Abraira, Pablo; Rodríguez Diéguez, Antonio; Melguizo Alonso, Consolación; Prados Salazar, José Carlos; Fernández, Ignacio A new family of chiral amino acid-based anthraquinones has been synthesized and fully characterized by multinuclear magnetic resonance (1H, 13C and 15N), elemental analysis, UV–Vis spectroscopy, circular dichroism, single-crystal X-ray diffraction and infrared spectroscopy. These molecules have been studied in different cancer cell lines (PANC-1, MCF-7, A549, SF268, HCT116) and a non-tumoral cell line (L-929) through different in vitro assays. The results showed that these molecules had dose-dependent antitumor effects, with compound 4e showing the lowest IC50 values across most tumor cell lines. Its crystal structure confirmed the enantiomeric purity and revealed π–π stacking interactions between anthraquinone and phenylalanine rings, features that may contribute to its enhanced biological performance. In addition, it was observed that the compounds had the ability to decrease cell migration and clonogenicity in the treated cell lines, demonstrating their ability to inhibit tumor malignancy processes. After studying possible molecular routes through which they induce their effect, it was shown that they are capable of inducing apoptosis and that they do not cause intracellular damage through the production of ROS. Therefore, these new synthesized compounds showed therapeutic potential in cancer, although further study of the molecular mechanisms involved in this antitumor activity is still needed.

Schatoff, Emma M Goswami, Sunkanya Zafra, Maria Paz Foronda, Miguel Shusterman, Michael Leach, Benjamin I Katti, Alyna Diaz, Bianca J Dow, Lukas E https://hdl.handle.net/10481/110845 2026-02-11T08:02:54Z

Distinct Colorectal Cancer–Associated APC Mutations Dictate Response to Tankyrase Inhibition Schatoff, Emma M; Goswami, Sunkanya; Zafra, Maria Paz; Foronda, Miguel; Shusterman, Michael; Leach, Benjamin I; Katti, Alyna; Diaz, Bianca J; Dow, Lukas E The majority of colorectal cancers show hyperactivated WNT signaling due to inactivating mutations in the adenomatous polyposis coli (APC) tumor suppressor. Genetically restoring APC suppresses WNT and induces rapid and sustained tumor regression, implying that reengaging this endogenous tumor-suppressive mechanism may be an effective therapeutic strategy. Here, using new animal models, human cell lines, and ex vivo organoid cultures, we show that tankyrase (TNKS) inhibition can control WNT hyperactivation and provide long-term tumor control in vivo, but that effective responses are critically dependent on how APC is disrupted. Mutant APC proteins truncated within the mutation cluster region physically engage the destruction complex and suppress the WNT transcriptional program, while APC variants with early truncations (e.g., ApcMin) show limited interaction with AXIN1 and β-catenin, and do not respond to TNKS blockade. Together, this work shows that TNKS inhibition, like APC restoration, can reestablish endogenous control of WNT/β-catenin signaling, but that APC genotype is a crucial determinant of this response. SIGNIFICANCE: This study reveals how subtle changes to the mutations in a critical colorectal tumor suppressor, APC, influence the cellular response to a targeted therapy. It underscores how investigating the specific genetic alterations that occur in human cancer can identify important biological mechanisms of drug response and resistance. This work was supported by a project grant from the NIH/NCI (CA195787-01) and a Stand Up To Cancer Colorectal Cancer Dream Team Translational Research Grant (SU2C-AACR-DT22-17). Stand Up To Cancer (SU2C) is a division of the Entertainment Industry Foundation. Research grants are administered by the American Association for Cancer Research, the scientific partner of SU2C. We thank Kevin Blighe for assistance with the Enhanced Volcano R package. We thank Shiaoching Gong and the MSKCC Mouse Transgenic Core Facility who performed zygote microinjections, supported in part by a U54 grant from the NIH/NCI (U54OD020355). We thank Katia Manova and Mesruh Turkekul from the Molecular Cytology Core Facility who performed ISH experiments, supported in part by a core grant from the NIH (P30 CA0088748). E.M. Schatoff was supported by a Medical Scientist Training Program grant from the National Institute of General Medical Sciences of the NIH under award number T32GM07739 to the Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program, and an F31 Award from the NCI/NIH under grant number 1 F31 CA224800-01. M.P. Zafra is supported in part by NCI grant NIH T32 CA203702. L.E. Dow was supported by a K22 Career Development Award from the NCI/NIH (CA 181280-01). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.

Han, Teng Goswami, Sukanya Hu, Yang Tang, Fanying Zafra, María Paz Murphy, Charles Cao, Zhen Poirier, John T. Khurana, Ekta Elemento, Olivier Hechtman, Jaclyn F. Ganesh, Karuna Yaeger, Rona Dow, Lukas E. https://hdl.handle.net/10481/110356 2026-01-27T12:44:10Z

Lineage Reversion Drives WNT Independence in Intestinal Cancer Han, Teng; Goswami, Sukanya; Hu, Yang; Tang, Fanying; Zafra, María Paz; Murphy, Charles; Cao, Zhen; Poirier, John T.; Khurana, Ekta; Elemento, Olivier; Hechtman, Jaclyn F.; Ganesh, Karuna; Yaeger, Rona; Dow, Lukas E. The WNT pathway is a fundamental regulator of intestinal homeostasis, and hyperactivation of WNT signaling is the major oncogenic driver in colorectal cancer. To date, there are no described mechanisms that bypass WNT dependence in intestinal tumors. Here, we show that although WNT suppression blocks tumor growth in most organoid and in vivo colorectal cancer models, the accumulation of colorectal cancer-associated genetic alterations enables drug resistance and WNT-independent growth. In intestinal epithelial cells harboring mutations in KRAS or BRAF, together with disruption of TP53 and SMAD4, transient TGFβ exposure drives YAP/TAZ-dependent transcriptional reprogramming and lineage reversion. Acquisition of embryonic intestinal identity is accompanied by a permanent loss of adult intestinal lineages, and long-term WNT-independent growth. This work identifies genetic and microenvironmental factors that drive WNT inhibitor resistance, defines a new mechanism for WNT-independent colorectal cancer growth, and reveals how integration of associated genetic alterations and extracellular signals can overcome lineage-dependent oncogenic programs. SIGNIFICANCE: Colorectal and intestinal cancers are driven by mutations in the WNT pathway, and drugs aimed at suppressing WNT signaling are in active clinical development. Our study identifies a mechanism of acquired resistance to WNT inhibition and highlights a potential strategy to target those drug-resistant cells This work was supported by a Research Scholar Award from the American Cancer Society (RSG-17-202-01-TBG), a project grant from the NIH/NCI (1R01CA222517-01A1), project grants from the Starr Cancer Consortium (#I10-0095 and #I11-0040) and a Stand Up To Cancer Colorectal Cancer Dream Team Translational Research Grant (SU2C-AACR-DT22-17). Research grants are administered by the American Association for Cancer Research, a scientific partner of SU2C. M.P. Zafra is supported in part by NCI grant NIH T32 CA203702.

Annunziato, Stefano Lutz, Catrin Henneman, Linda Bhin, Jinhyuk Wong, Kim Siteur, Bjørn Gerwen, Bas van de Korte-Grimmerink, Renske Zafra, Maria Paz Schatoff, Emma M Drenth, Anne Paulien van der Burg, Eline Eijkman, Timo Mukherjee1, Siddhartha2 Boroviak, Katharina Wessels, Lodewyk FA van de Ven, Marieke Huijbers, Ivo J Adams, David J Dow, Lukas E Jonkers, Jos https://hdl.handle.net/10481/110328 2026-01-27T11:05:35Z

In situ CRISPR-Cas9 base editing for the development of genetically engineered mouse models of breast cancer Annunziato, Stefano; Lutz, Catrin; Henneman, Linda; Bhin, Jinhyuk; Wong, Kim; Siteur, Bjørn; Gerwen, Bas van; de Korte-Grimmerink, Renske; Zafra, Maria Paz; Schatoff, Emma M; Drenth, Anne Paulien; van der Burg, Eline; Eijkman, Timo; Mukherjee1, Siddhartha2; Boroviak, Katharina; Wessels, Lodewyk FA; van de Ven, Marieke; Huijbers, Ivo J; Adams, David J; Dow, Lukas E; Jonkers, Jos Genetically engineered mouse models (GEMMs) of cancer have proven to be of great value for basic and translational research. Although CRISPR-based gene disruption offers a fast-track approach for perturbing gene function and circumvents certain limitations of standard GEMM development, it does not provide a flexible platform for recapitulating clinically relevant missense mutations in vivo. To this end, we generated knock-in mice with Cre-conditional expression of a cytidine base editor and tested their utility for precise somatic engineering of missense mutations in key cancer drivers. Upon intraductal delivery of sgRNA-encoding vectors, we could install point mutations with high efficiency in one or multiple endogenous genes in situ and assess the effect of defined allelic variants on mammary tumorigenesis. While the system also produces bystander insertions and deletions that can stochastically be selected for when targeting a tumor suppressor gene, we could effectively recapitulate oncogenic nonsense mutations. We successfully applied this system in a model of triple-negative breast cancer, providing the proof of concept for extending this flexible somatic base editing platform to other tissues and tumor types.

Katti, Alyna Foronda, Miguel Zimmerman, Jill Diaz, Bianca Zafra, Maria Paz Goswami, Sukanya Dow, Lukas E. https://hdl.handle.net/10481/110326 2026-01-27T11:03:50Z

GO: a functional reporter system to identify and enrich base editing activity Katti, Alyna; Foronda, Miguel; Zimmerman, Jill; Diaz, Bianca; Zafra, Maria Paz; Goswami, Sukanya; Dow, Lukas E. Base editing (BE) is a powerful tool for engineering single nucleotide variants (SNVs) and has been used to create targeted mutations in cell lines, organoids and animal models. Recent development of new BE enzymes has provided an extensive toolkit for genome modification; however, identifying and isolating edited cells for analysis has proven challenging. Here we report a 'Gene On' (GO) reporter system that indicates precise cytosine or adenine base editing in situ with high sensitivity and specificity. We test GO using an activatable GFP and use it to measure the kinetics, efficiency and PAM specificity of a range of new BE variants. Further, GO is flexible and can be easily adapted to induce expression of numerous genetically encoded markers, antibiotic resistance genes or enzymes, such as Cre recombinase. With these tools, GO can be exploited to functionally link BE events at endogenous genomic loci to cellular enzymatic activities in human and mouse cell lines and organoids. Thus, GO provides a powerful approach to increase the practicality and feasibility of implementing CRISPR BE in biomedical research.