bioRxiv Science⌕ Search

Biology subjects

Hautaniemi, S.

Publications and source records attributed to Hautaniemi, S..

At least 19 recordsLinked to original sources

FEATMAP: Targeted Correction of Acquisition Signatures Harmonizes Medical Foundation Model Embeddings and Enables Robust Task Generalization

AO_SCPLOWBSTRACTC_SCPLOWMedical foundation models compress biomedical data into embeddings that support diverse downstream clinical tasks. However, successful model deployment is hampered by performance degradation on external data. It is recognized that embeddings capture acquisition signatures, such as hardware and technical differences, in addition to biology. Effective harmonization must remove the acquisition signature while preserving biological signals, a trade-off that current methods fail to balance adequately. Input-level normalization fails to eliminate acquisition signatures from embeddings, whereas embedding-level methods adjust features in an untargeted manner. We present FEATMAP, a harmonization approach that models acquisition signatures as geometric distortions between manifolds of similarly arranged embeddings. Using paired data that isolate the effect of acquisition signatures, FEATMAP fits a single global affine transformation per foundation model to correct acquisition signatures directly in the embedding space. This targeted, reusable correction aims to preserve biological and demographic variation while harmonizing across acquisition signatures. Across scanner and foundation-model harmonization in digital pathology and field-strength harmonization in brain MRI, FEATMAP improves cross-condition embedding similarity, reduces performance gaps without retraining, and suggests potential for the alignment of disparate embedding spaces.

bioinformatics↗

Principles of subclonal gene dosage across human cancer

Intratumor heterogeneity drives disease progression, but even though subclonal copy number variation (CNV) is a major contributor to this heterogeneity1, its impact on cell phenotype is not fully understood. Here, by applying high-quality joint whole genome sequencing and mRNA profiling in single cells (DNTR-seq2) to solid tumors and leukemias from 57 patients, we have analyzed the in vivo transcriptional effect of subclonal CNVs within a tumor. We found that gene dosage is generally additive in low and moderate copy states, but that cancer-type-specific compensation is common, and core promoter elements are associated with reduced additivity. We find that different classes of subclonal CNV impose varying degrees of transcriptional constraints on a cell, with highly amplified megabase-size regions associated with a strong effect both in cis and trans, while arm level CNVs generally exert a milder effect. We also describe a previously unappreciated class of tumors with transient clonality, where every cell is genetically highly distinct. We find that transient clonality is common in ovarian cancer and soft tissue sarcoma, that it is preceded by a whole genome duplication event, and that gene dosage in these tumors affects transcript abundance to a similar degree as in cancer with stable subclones.

cancer biology↗

Single-Cell Transcriptomic Signatures Enable Stratified Combination Therapy for Platinum-Resistant Ovarian Cancer

In high-grade serous carcinoma (HGSC), extensive intra-tumoral heterogeneity hinders complete eradication and remains a major obstacle to developing combination therapies capable of eliminating subpopulations resistant to standard-of-care treatment. Using single-cell RNA sequencing of 72 samples from 54 HGSC patients spanning treatment-naive, post-neoadjuvant chemotherapy and relapse stages, we established a carboplatin-anchored framework that identifies transcriptional signatures of intrinsic (pre-existing) and adaptive (therapy-induced) resistance in individual tumors and prioritizes mechanistically matched drugs to potentiate carboplatin efficacy. Candidate compounds were ranked by integrating orthogonal resources--viability (GDSC, PRISM) and perturbational transcriptomics (L1000, Perturb-seq)--to reduce context bias. Among 64 candidates, three carboplatin adjuvants enhanced long-term efficacy in patient-derived organoids (PDOs), and pevonedistat further significantly reduced tumor burden in orthotopic xenografts. This tiered validation pipeline--from short-term and long-term PDOs and in vivo orthoptic xenografts--establishes a translational framework linking single cell resistance programs to actionable, tumor-specific, carboplatin-anchored combinations for HGSC.

cancer biology↗

Histolytics: A Panoptic Spatial Analysis Framework for Interpretable Histopathology

Quantifying spatial organization in hematoxylin and eosin (H&E)-stained whole-slide images (WSIs) is essential for uncovering tissue-level patterns relevant to pathology. We present Histolytics, an open-source, scalable Python framework for interpretable, WSI-scale histopathological analysis. Histolytics integrates panoptic segmentation with spatial querying, morphological profiling, and graph-based analytics to enable high-resolution, quantitative characterization of nuclei, tissue compartments, and the extracellular matrix (ECM). Designed to align with diagnostic reasoning, Histolytics supports segmentation with state-of-the-art deep learning models and provides modular tools for extracting biologically grounded features across entire WSIs. By leveraging spatially contextualized measurements at cellular and tissue levels, Histolytics addresses a critical gap in explainable computational pathology, offering an interpretable alternative or complement to black-box predictive models. The framework is compatible with the broader Python data science ecosystem and includes extensive documentation and pretrained models to promote widespread adoption.

bioinformatics↗

Multi-modal characterization of transcriptional programs that drive metastatic cascades to solid sites and ascites in ovarian cancer

Ovarian high-grade serous carcinoma (HGSC) is characterized by extensive intra-peritoneal dissemination and tumor heterogeneity. In the metastatic cascade, tumors utilize several transcriptional programs to translocate and survive in distant tissues. Here, we analyzed multi-modal, real-world data from 350 tumor samples across 160 patients with HGSC to identify transcriptional programs that drive intra-peritoneal metastasis and heterogeneity. We identified nine transcriptional programs, including those regulating epithelial-mesenchymal transition and immune modulation and cytoskeletal reorganization, which shape distinct metastatic trajectories to solid and ascitic environments and are associated to treatment response. Our results reveal pronounced intra-patient transcriptional heterogeneity, which in some cases surpassed inter-patient heterogeneity, highlighting the importance of multi-site sampling for accurate prognostication and combinatorial treatments in HGSC. Our extensive characterization offers novel insights into intra-peritoneal metastasis with significant prognostic implications, reveals histomorphological biomarkers for patient stratification and paves the way for innovative therapeutic strategies aimed at impairing cancer cell adaptability and limiting metastasis.

cancer biology↗

Proteome profiling reveals HES1-driven mitotic catastrophe in ovarian serous carcinoma

Ovarian high-grade serous cancer (HGSC) is an aggressive subtype of epithelial ovarian cancer. Here, we identify BX-912, a phosphoinositide-dependent kinase 1 (PDPK1) inhibitor, as a promising therapeutic agent for HGSC. BX-912 suppressed HGSC growth as a single agent and synergized with olaparib independently of BRCA status. Unexpectedly, BX-912 treatment induced multinucleation, a phenotype not observed with other PDPK1 inhibitors. Proteome Integral Solubility Alteration (PISA) profiling revealed the transcription factor HES1 as a functional target of BX-912. Structural modeling showed that BX-912 binds the Orange domain of HES1, while its WRPW motif mediates interactions with protein partners, including the AP2 endocytic protein complex, coordinating their nuclear accumulation that leads to a mitotic catastrophe. Furthermore, cell cycle analyses showed that BX-912 combined with olaparib synergistically enhanced DNA damage and G2-M arrest. Our study demonstrates the value of proteomics for revealing hidden drug activities. It also identifies potential inhibition strategies for HES1, which is commonly overexpressed in HGSC. Additionally, this study proposes a novel strategy of targeting consecutive cell cycle phases to enhance treatment efficacy in HGSC.

cancer biology↗

Multi-Omics Characterization of ctDNA Release Mechanisms in Ovarian Cancer

Circulating tumor DNA (ctDNA) is a powerful biomarker capable to predict tumor dynamics and treatment response. Despite its importance, the biological mechanisms behind ctDNA release have remained unclear. The variability among patients is affected by cancer burden, histology and stage, but these factors cover only a part of the detected variability. Herein, we characterized molecular drivers of baseline ctDNA variability in a real-world cohort of 118 patients with ovarian high-grade serous carcinoma (HGSC). Genomic and transcriptomic analyses revealed a strong positive correlation between ctDNA levels and cellular proliferation, and an inverse relationship with immune activity. Particularly, low ctDNA tumors exhibited higher bulk expression of mucins and CIITA, suggesting that intrinsic immune responses and immune landscape are linked to ctDNA release. The low and high ctDNA levels were significantly associated with poor prognosis compared to medium level in unresectable HGSC patients treated with neoadjuvant chemotherapy. In summary, our results suggest that ctDNA release is modulated by cancer cell proliferation and tumor microenvironment.

cancer biology↗

SegmentQTL: Identifying genetic variants influencing molecular phenotypes in copy number-driven cancers

MotivationMolecular quantitative trait loci (molQTL) analysis links genetic variants to molecular phenotypes, such as gene expression, but existing tools do not account for the structural complexity of copy number-driven cancers. High genomic instability of these cancers leads to chromosomal breaks (breakpoints), which disrupt the physical connection between genes and adjacent regulatory elements. Standard molQTL methods are unable to accommodate breakpoint information and would therefore indiscriminately test associations across breakpoints, leading to spurious signals and reduced biological relevance. To address these challenges, we developed SegmentQTL, a segmentation-aware molQTL analysis tool, designed to improve the accuracy of association testing in unstable cancer genomes by incorporating sample-specific break-point information. ResultsSegmentQTL applies an integrated purifying filtering step that removes associations spanning breakpoints, ensuring that only variants within the same segment as the phenotype are tested. This prevents false discoveries and reduces background noise. We evaluated SegmentQTL on selected genes from stable and unstable genomic regions and compared its results with a previously published state-of-the-art tool. In stable regions, SegmentQTL produced similar results, validating its approach. In unstable regions, however, the filtering step refined detected associations by shifting peak locations and removing artefactual signals that would arise if genomic instability were not properly accounted for. Availability and implementationhttps://github.com/HautaniemiLab/SegmentQTL

bioinformatics↗

Identification of Monotonically Classifying Pairs of Genes for Ordinal Disease Outcomes

In this study, we extend an existing classification method for identifying pairs of genes whose joint expression is associated with binary outcomes to ordinal multi-class outcomes, such as overall survival or disease progression. Our approach is motivated by the need for interpretable classifiers that can provide insights into the underlying biological mechanisms. It can be easily adapted to different research questions, such as identifying gene pair signatures or functional enrichment. We demonstrate that our method is comparable to state-of-the-art classification approaches in terms of performance, while offering the benefit of higher interpretability and adaptability to solve different research questions. Our evaluation on two real-world use cases in glioblastoma and high-grade serous ovarian carcinoma shows that our approach can effectively predict ordinal outcomes and provide novel biological insights. The code is available at https://github.com/oceanefrqt/MBMC.

systems biology↗

Multi-Omics Analysis Reveals the Attenuation of the Interferon Pathway as a Driver of Chemo-Refractory Ovarian Cancer

Ovarian high-grade serous carcinoma (HGSC) represents the deadliest gynecological malignancy, with 10-15% of patients exhibiting primary resistance to first-line chemotherapy. These primarily chemo-refractory patients have particularly poor survival outcomes, emphasizing the urgent need for developing predictive biomarkers and novel therapeutic approaches. Here, we show that interferon type I (IFN-I) pathway activity in cancer cells is a crucial determinant of chemotherapy response in HGSC. Through a comprehensive multi-omics analysis within the DECIDER observational trial (ClinicalTrials.gov identifier NCT04846933) cohort, we identified that chemo-refractory HGSC is characterized by diminished IFN-I and enhanced hypoxia pathway activities. Importantly, IFN-I pathway activity was independently prognostic for patient survival, highlighting its potential as a biomarker. Our results elucidate the heterogeneity of treatment response at the molecular level and suggest that augmentation of IFN-I response could enhance chemosensitivity in refractory cases. This study underscores the potential of the IFN-I pathway as a therapeutic target and advocates for the initiation of clinical trials testing external modulators of the IFN-I response, promising a significant stride forward in the treatment of refractory HGSC.

cancer biology↗

Chemotherapy induces myeloid-driven spatial T-cell exhaustion in ovarian cancer

To uncover the intricate, chemotherapy-induced spatiotemporal remodeling of the tumor microenvironment, we conducted integrative spatial and molecular characterization of 97 high-grade serous ovarian cancer (HGSC) samples collected before and after chemotherapy. Using single-cell and spatial analyses, we identify increasingly versatile immune cell states, which form spatiotemporally dynamic microcommunities at the tumor-stroma interface. We demonstrate that chemotherapy triggers spatial redistribution and exhaustion of CD8+ T cells due to prolonged antigen presentation by macrophages, both within interconnected myeloid networks termed "Myelonets" and at the tumor stroma interface. Single-cell and spatial transcriptomics identifies prominent TIGIT-NECTIN2 ligand-receptor interactions induced by chemotherapy. Using a functional patient-derived immuno-oncology platform, we show that CD8+T-cell activity can be boosted by combining immune checkpoint blockade with chemotherapy. Our discovery of chemotherapy-induced myeloid-driven spatial T-cell exhaustion paves the way for novel immunotherapeutic strategies to unleash CD8+ T-cell-mediated anti-tumor immunity in HGSC.

cancer biology↗

Comparative Analysis of Gene Expression Analysis Methods for RNA In Situ Hybridization Images

Gene expression analysis is pivotal in cancer research and clinical practice. While traditional methods lack spatial context, RNA in situ hybridization (RNA-ISH) is a powerful technique that retains spatial tissue information. Here, we investigated RNAscope score, RT-droplet digital PCR (RT-ddPCR), and automated QuantISH and QuPath in quantifying RNA-ISH expression values from formalin-fixed paraffin-embedded samples. We compared the methods using high-grade serous ovarian carcinoma samples, focusing on CCNE1, WFDC2, and PPIB genes. Our findings demonstrate good concordance between automated methods and RNAscope, with RT-ddPCR showing less concordance. We conclude that QuantISH exhibits robust performance, even for low-expressed genes like CCNE1, showcasing its modular design and enhancing accessibility as a viable alternative for gene expression analysis.

bioinformatics↗

Patient-derived functional immuno-oncology platform identifies responders to ATR inhibitor and immunotherapy combinations in ovarian cancer

Responses to single agent immunotherapies have remained modest in high-grade serous ovarian cancer (HGSC), suggesting the need for combination treatments. Identifying clinically effective immunotherapy combinations (IC) requires pre-clinical testing using models representing the patient-specific immune microenvironment. Here, we established a functional immuno-oncology platform for high-throughput and functional testing of IC using HGSC patient-derived immunocompetent cultures (iPDCs) established on patient-derived omentum gel matrix. We employed genomic and single-cell analysis to assess the intricate and functional characteristics of the iPDCs combined with tumor and immune cell-specific cytotoxic responses. Corroborating the clinical response to Poly (ADP-ribose) polymerase inhibitors (PARPi), iPDCs showed homologous recombination deficiency (HRD) - specific response to PARPi. Importantly, drug responses from iPDCs of chemotherapy and PARPi refractory patients corresponded with patient outcomes and aligned with distinct pathway activities from single-cell RNA sequencing analysis. Furthermore, iPDCs from HRD tumors showed response to anti-PD1 antibody as measured by decrease in tumor cells combined with augmented T cell activation. High-throughput drug testing followed by single cell-imaging from iPDCs revealed patient-specific responses to combination of ataxia telangiectasia and Rad3-related inhibitor (ATRi) with DNA damaging agents or immunotherapies. Integration of cytotoxic responses with immune cell states uncovered patient-specific immune activation with the combination of ATRi and a novel immunotherapy targeting Autotaxin (ATX), and this response was significantly associated with a tumor-cell replication stress biomarker in single-cell analysis of tCycIF highly multiplexed imaging. In conclusion, iPDCs provide a platform for high-throughput screening and functional testing of immuno-oncology agents for precision oncology in HGSC.

cancer biology↗

Deciphering Cancer Genomes with GenomeSpy: A Grammar-Based Visualization Toolkit

BackgroundVisualization is an indispensable facet of genomic data analysis. Despite the abundance of specialized visualization tools, there remains a distinct need for tailored solutions. However, their implementation typically requires extensive programming expertise from bioinformaticians and software developers, especially when building interactive applications. Toolkits based on visualization grammars offer a more accessible, declarative way to author new visualizations. Nevertheless, current grammar-based solutions fall short in adequately supporting the interactive analysis of large data sets with extensive sample collections, a pivotal task often encountered in cancer research. ResultsWe present GenomeSpy, a grammar-based toolkit for authoring tailored, interactive visualizations for genomic data analysis. Users can implement new visualization designs with little effort by using combinatorial building blocks that are put together with a declarative language. These fully customizable visualizations can be embedded in web pages or end-user-oriented applications. The toolkit also includes a fully customizable but user-friendly application for analyzing sample collections, which may comprise genomic and clinical data. Findings can be bookmarked and shared as links that incorporate provenance information. A distinctive element of GenomeSpys architecture is its effective use of the graphics processing unit (GPU) in all rendering. GPU usage enables a high frame rate and smoothly animated interactions, such as navigation within a genome. We demonstrate the utility of GenomeSpy by characterizing the genomic landscape of 753 ovarian cancer samples from patients in the DECIDER clinical trial. Our results expand the understanding of the genomic architecture in ovarian cancer, particularly the diversity of chromosomal instability. We also show how GenomeSpy enabled the discovery of clinically actionable genomic aberrations. ConclusionsGenomeSpy is a visualization toolkit applicable to a wide range of tasks pertinent to genome analysis. It offers high flexibility and exceptional performance in interactive analysis. The toolkit is open source with an MIT license, implemented in JavaScript, and available at https://genomespy.app/.

bioinformatics↗

Direct conversion of human fibroblasts to pancreatic epithelial cells through transient progenitor states is controlled by temporal activation of defined factors

Cell fate can be reprogrammed by ectopic expression of lineage-specific transcription factors (TF). For example, few specialized cell types like neurons, hepatocytes and cardiomyocytes have been generated from fibroblasts by defined factors (Wang et al, 2021). However, the exact cell state transitions and their control mechanisms during cell fate conversion are still poorly understood. Moreover, the defined TFs for generating vast majority of the human cell types are still elusive. Here, we report a novel protocol for reprogramming human fibroblasts to pancreatic exocrine cells with phenotypic and functional characteristics of ductal epithelial cells using a minimal set of six TFs. We mapped the molecular determinants of lineage dynamics at single-cell resolution using a novel factor-indexing method based on single-nuclei multiome sequencing (FI-snMultiome-seq) that enables dissecting the role of each individual TF and pool of TFs in cell fate conversion. We show that transdifferentiation - although being considered a direct cell fate conversion method - occurs through transient progenitor states orchestrated by stepwise activation of distinct TFs. Specifically, transition from mesenchymal fibroblast identity to epithelial pancreatic exocrine fate involves two deterministic steps: first, an endodermal progenitor state defined by activation of HHEX concurrently with FOXA2 and SOX17, and second, temporal GATA4 activation essential for maintenance of pancreatic cell fate program. Collectively, our data provide a high-resolution temporal map of the epigenome and transcriptome remodeling events that facilitate cell fate conversion, suggesting that direct transdifferentiation process occurs through transient dedifferentiation to progenitor cell states controlled by defined TFs.

cell biology↗

A synthetic lethal dependency on casein kinase 2 in response to replication-perturbing drugs in RB1-deficient ovarian and breast cancer cells

Treatment of patients with high-grade serous ovarian carcinoma (HGSOC) and triple-negative breast cancer (TNBC) includes platinum-based drugs, gemcitabine, and PARP inhibitors. However, resistance to these therapies develops in most cases, highlighting the need for novel therapeutic approaches and biomarkers to guide the optimal treatment choice. Using a CRISPR loss-of-function screen for carboplatin sensitizers in the HGSOC cell line OVCAR8, we identified CSNK2A2, the gene encoding for the alpha () catalytic subunit of casein kinase 2 (CK2). Expanding on this finding, we confirmed that the CK2 inhibitors silmitasertib and SGC-CK2-1 sensitized many, but not all, TNBC and HGSOC cell lines to the drugs that perturb DNA replication, including platinum drugs, gemcitabine, and PARP inhibitors. We identified RB1 tumor suppressor deficiency as a prerequisite context for the CK2 inhibition-mediated sensitization to these therapeutics. In RB1-deficient cells, CK2 inhibition resulted in accumulation of cells in S phase of the cell cycle, associated with micronuclei formation, and accelerated PARP inhibitor-induced aneuploidy and mitotic cell death. Patient HGSOC organoids that lacked RB1 expression displayed an enhanced long-term response to carboplatin and PARP inhibitor niraparib when combined with silmitasertib, suggesting RB1-stratified efficacy in patients. As RB1 deficiency affects up to 25% of HGSOC and 40% of TNBC cases, CK2 inhibition, proven safe from previous clinical exploration with silmitasertib, is a promising approach to overcome resistance to standard therapeutics in large strata of patients.

cancer biology↗

Lineage-specific oncogenes drive growth of major forms of human cancer using common downstream mechanisms

Mutations in hundreds of genes have been associated with formation of human cancer, with different oncogenic lesions prevalent in different cancer types. Yet, the malignant phenotype is simple, characterized by unrestricted growth of cells that invade neighboring healthy tissue and in many cases metastasize to distant organs. One possible hypothesis explaining this dichotomy is that the cancer genes regulate a common set of target genes, which then function as master regulators of essential cancer phenotypes, such as growth, invasion and metastasis. To identify mechanisms that drive the most fundamental feature shared by all tumors - unrestricted cell proliferation - we used a multiomic approach to identify common transcriptional and posttranslational targets of major oncogenic pathways active in different cancer types, and combined this analysis with known regulators of the cell cycle. We identified translation and ribosome biogenesis as common targets of both transcriptional and posttranslational oncogenic pathways. By combining proteomic analysis of clinical samples with functional studies of cell cultures, we also establish NOLC1 as a key node whose convergent regulation both at transcriptional and posttranslational level is critical for tumor cell proliferation. Our results indicate that lineage-specific oncogenic pathways commonly regulate the same set of targets important for growth control, revealing novel key downstream nodes that could be targeted for cancer therapy or chemoprevention.

cancer biology↗

L1 retrotransposition is regulated post-transcriptionally In High-Grade Serous Ovarian Cancer

L1 retrotransposons are the only protein-coding active transposable elements in the human genome. Although silenced during normal conditions, they are highly expressed in human epithelial cancers including high-grade serous ovarian cancer (HGSC), where they transcribe to form L1 mRNA and subsequently integrate into the genome by a process called retrotransposition. Despite of high L1 protein expression in the earliest phases of HGSC, these tumors do not accrue many somatic L1 insertions. To understand this unexplained disconnect, we monitored the transcription and retrotransposition activity of two frequently expressed retrotransposition-competent (RC)-L1 (RC-L1) in 64 clinical tumor specimens from 34 HGSC patients and found that despite the presence of RC-L1 mRNA, a third of samples did not acquire somatic L1 insertions. In addition to high inter-patient variability in retrotransposition frequency, there was remarkable intra-patient heterogeneity in L1 insertion patterns between tumor sites, indicating that L1 retrotransposition is highly dynamic in vivo. Comparison of genomic and transcriptomic features of L1-null tumors with L1-high tumors (those with [≥]5 somatic L1 insertions) showed that retrotransposition was favored by increased rate of cell proliferation.

cancer biology↗