Distinctive phenogroup to differentiate diagnosis of cardiac myxoma vs cardiovascular disease examining blood-based circulating cell biomarkers

Figures

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  Figure 1. Experimental workflow. Cardiac myxoma (CM) tissue biopsies were processed pooled and stained distinguishing central and peripheral tumor regions. Primary cell cultures enriched for cancer cells were obtained from tumor tissue and peripheral blood of CM patients. Phenotypic profile of cancer and endothelial cells were analyzed by immunological testing in tissue, cell cultures and PDXs. PDX patient derived xenograft, IHC immunohistochemistry, CTCs circulating tumor cells, ICC immunocytochemistry, CECs circulating endothelial cells.

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  Figure 2. Cancer and endothelial cell characterization in tumor biopsy. Tumor biopsy was processed to distinguish central and peripheral regions of the cardiac myxoma. Immunohistochemical analysis for the expression of calretinin shown a different distribution of calretinin positivity cells prevalently distributed in the central region of the tumors (P = 0.01) and for CD31 expression (P = 0.001). Primary culture tumor region derived, preserved calretinin positive cells and showing at FACS analysis a different distribution of cell CD45 negative CD34 positive (P = 0.008). PDXs derived distinguishing central and peripheral region confirmed the prevalent density of vessel-forming units in the central area of cardiac myxoma.

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  Figure 3. Cancer and endothelial profiles in cardiac myxoma liquid biopsy. Immunocytochemistry analysis shows the presence of cells calretinin positive (also visible at immunofluorescence assay) and CD31 positive. Cells CD31 positive are recognizable in blood-derived cultures of cardiovascular patients and healthy subject as expression of vessel forming unit cells prevalently presented in the peripheral blood of cardiovascular rather that CM patients.

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  Figure 4. Visual graphical illustration of phenogroup blood-based cellular biomarkers. (A) The exchange of mediators and cells between the heart and the systemic circulation affects the type of non-hematological cells within the peripheral vessel (B) characterizing the cell type observed by blood collection (C) the graph shows the distribution of the circulating endothelial cells subsets in function of the subset of endothelial cells isolated from different region of tissue biopsy. (D) In the healthy subject the condition of heart health determines a profile mainly characterized by the CD34 bright cells which are part of the progenitor compartment with multiple differentiation abilities. In (E) the heart disease patients the pathological condition of the heart determines a greater recruitment of committed progenitors in the endothelial sense probably to support the increased demand of blood flow. (F) In cardiac myxoma the heart is the site of a proliferative lesion which autonomously contributes to the non-hematological component of the blood, evidencing a tissue remodeling which also includes the intratumor vascular network.

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  Figure 5. Blood- based cellular biomarkers of other cardiac tumors. (A) Histopathological section (magnification 20 ×) of the tumor stained with hematoxylin and eosin is reported of a case of atrial villous angiomyolipoma, diagnosed in a 39-year-old male. In the left top of the image its echocardiographic picture shows the mass arising from the atrial wall. In the right bottom of the image, a particular of a tumor cell (magnification 40 ×). In (B) his cultured circulating cells specimen (magnification 10 ×) shows in the right bottom of the image, cytomorphological feature like tumor cell in primary lesion (magnification 40 ×). (C) Histopathological section (magnification 20 ×) of a case of atrial myxoma, hematoxylin and eosin stained, diagnosed in a 60-year-old male during an echocardiographic control. Echocardiographic picture is reported in the left top of the image and in (D) cultured circulating cells (magnification 20 ×), in the right bottom of the image a particular of a calretinin positive cell (magnification 40 ×). (E) and (F) show cultured circulating cells isolated from blood sample of a 20-year-old female angiosarcoma patient and a case of 69-year-old male fibroelastoma,…