RGD Collagen for Engineering a Contractile Tissue and Cell Therapy after Myocardial Infarct

Currently, the clinical impact of cell therapy after a myocardial infarction (MI) is limited by low cell engraftment due to significant cell death, including apoptosis, in an infarcted, inflammatory, poor angiogenic environment, low cell retention and secondary migration. Cells interact with their environment through integrin mechanoreceptors that control their survival/apoptosis/differentiation/migration/proliferation. Optimizing these interactions may be a way of improving outcomes. The association of free cells with a 3D-scaffold may be a way to target their integrins. Collagen is the most abundant structural component of the extracellular matrix (ECM) and the best contractility levels are achieved with cellular preparations containing collagen, fibrin, or Matrigel (i.e. tumor extract). In the interactions between cells and ECM, 3 main proteins are recognised: collagen, laminin and RGD (Arg-Gly-Asp) peptide. The RGD plays a key role in heart development, after MI, and on cardiac cells. Cardiomyocytes secrete their own laminin on collagen. The collagen has a non-functional cryptic RGD and is thus suboptimal for interactions with associated cells. The use of a collagen functionalized with RGD may help to improve collagen biofunctionality. It may help in the delivery of paracrine cells, whether or not they are contractile, and in assisting tissue engineering a safe contractile tissue.

Evaluation of the potential of Pap test fluid and cervical swabs to serve as clinical diagnostic biospecimens for the detection of ovarian cancer by mass spectrometry-based proteomics

Background The purpose of this study was to determine whether the residual fixative from a liquid-based Pap test or a swab of the cervix contained proteins that were also found in the primary tumor of a woman with high grade serous ovarian cancer. This study is the first step in determining the feasibility of using the liquid-based Pap test or a cervical swab for the detection of ovarian cancer protein biomarkers. Methods Proteins were concentrated by acetone precipitation from the cell-free supernatant of the liquid-based Pap test fixative or eluted from the cervical swab. Protein was also extracted from the patient’s tumor tissue. The protein samples were digested into peptides with trypsin, then the peptides were run on 2D-liquid chromatography mass spectrometry (2D-LCMS). The data was searched against a human protein database for the identification of peptides and proteins in each biospecimen. The proteins that were identified were classified for cellular localization and molecular function by bioinformatics integration. Results We identified almost 5000 proteins total in the three matched biospecimens. More than 2000 proteins were expressed in each of the three biospecimens, including several known ovarian cancer biomarkers such as CA125, HE4, and mesothelin. By Scaffold analysis of the protein Gene Ontology categories and functional analysis using PANTHER, the proteins were classified by cellular localization and molecular function, demonstrating that the Pap test fluid and cervical swab proteins are similar to each other, and also to the tumor extract. Conclusions Our results suggest that Pap test fixatives and cervical swabs are a rich source of tumor-specific biomarkers for ovarian cancer, which could be developed as a test for ovarian cancer detection.

Transforming the spleen into a liver-like organ in vivo

Regenerating human organs remains an unmet medical challenge. Suitable transplants are scarce, while engineered tissues have a long way to go toward clinical use. Here, we demonstrate a different strategy that successfully transformed an existing, functionally dispensable organ to regenerate another functionally vital one in the body. Specifically, we injected a tumor extract into the mouse spleen to remodel its tissue structure into an immunosuppressive and proregenerative microenvironment. We implanted autologous, allogeneic, or xenogeneic liver cells (either primary or immortalized), which survived and proliferated in the remodeled spleen, without exerting adverse responses. Notably, the allografted primary liver cells exerted typical hepatic functions to rescue the host mice from severe liver damages including 90% hepatectomy. Our approach shows its competence in overcoming the key challenges in tissue regeneration, including insufficient transplants, immune rejection, and poor vascularization. It may be ready for translation into new therapies to regenerate large, complex human tissue/organs.

Antimitotic activity of EA21b mammary-carcinoma extract

Many tumors produce factors that affect cell-cycle and cell proliferation. In the present study we have analyzed the effect of a mammary-tumor extract injection on the mitotic activity of several organs in young male C3H/S mice previously standardized for circadian periodicity. One-half of the animals received an intraperitoneal EA21b tumor extract dose at 16:00 h, while the other half received saline. Animals were sacrificed on the following day at 08:00, 12:00 or 16:00 h. 4 h after receiving an injection of colchicine by the same route. Samples of duodenum, kidney, liver, and submaxillary gland were excised and processed for hematoxylin-eosin staining. Mitotic indices, expressed as the number of colchicine-arrested metaphases per 1,000 nuclei, were assessed in convoluted tubule epithelium, duodenal crypt enterocytes, hepatocytes and submaxillary gland ductal and acinar sialocytes. All values were expressed as mean ± SEM. Statistical analyses were performed by ANOVA, Bonferroni and Student’s t-tests. In contrast to the mitotic indices reductions observed in renal convoluted tubules cells and duodenal crypt enterocytes, neither the submaxillary gland nor the liver were found to contain cell types whose mitotic activity was affected by the tumor extract. We conclude that EA21b mammary carcinoma contains one or more factors that inhibit the proliferation of selected populations of normal cells.