RESUMO
Genome sequencing of cancers often reveals mosaics of different subclones present in the same tumour1-3. Although these are believed to arise according to the principles of somatic evolution, the exact spatial growth patterns and underlying mechanisms remain elusive4,5. Here, to address this need, we developed a workflow that generates detailed quantitative maps of genetic subclone composition across whole-tumour sections. These provide the basis for studying clonal growth patterns, and the histological characteristics, microanatomy and microenvironmental composition of each clone. The approach rests on whole-genome sequencing, followed by highly multiplexed base-specific in situ sequencing, single-cell resolved transcriptomics and dedicated algorithms to link these layers. Applying the base-specific in situ sequencing workflow to eight tissue sections from two multifocal primary breast cancers revealed intricate subclonal growth patterns that were validated by microdissection. In a case of ductal carcinoma in situ, polyclonal neoplastic expansions occurred at the macroscopic scale but segregated within microanatomical structures. Across the stages of ductal carcinoma in situ, invasive cancer and lymph node metastasis, subclone territories are shown to exhibit distinct transcriptional and histological features and cellular microenvironments. These results provide examples of the benefits afforded by spatial genomics for deciphering the mechanisms underlying cancer evolution and microenvironmental ecology.
Assuntos
Neoplasias da Mama , Carcinoma Intraductal não Infiltrante , Evolução Clonal , Células Clonais , Genômica , Feminino , Humanos , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Carcinoma Intraductal não Infiltrante/genética , Carcinoma Intraductal não Infiltrante/patologia , Evolução Clonal/genética , Células Clonais/metabolismo , Células Clonais/patologia , Mutação , Microambiente Tumoral/genética , Sequenciamento Completo do Genoma , Transcriptoma , Reprodutibilidade dos Testes , Microdissecção , AlgoritmosRESUMO
We introduce a framework for the reconstruction of the amplitude, phase, and polarization of an optical vector-field using measurements acquired by an imaging device characterized by an integral transform with an unknown spatially variant kernel. By incorporating effective regularization terms, this new approach is able to recover an optical vector-field with respect to an arbitrary representation system, which may be different from the one used for device calibration. In particular, it enables the recovery of an optical vector-field with respect to a Fourier basis, which is shown to yield indicative features of increased scattering associated with tissue abnormalities. We demonstrate the effectiveness of our approach using synthetic holographic images and biological tissue samples in an experimental setting, where the measurements of an optical vector-field are acquired by a multicore fiber endoscope, and observe that indeed the recovered Fourier coefficients are useful in distinguishing healthy tissues from tumors in early stages of oesophageal cancer.
