PMC9772982_01
(A) The hands of normal and acromegaly people. (B) Preoperative MRI, 44mmx 27mmx37mm in size, with left cavernous sinus involvement, and internal carotid artery encased. (C) Postoperative MRI, show the residual tumor encircling the left internal carotid artery.
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The patient was a 20-year-old Chinese male who developed symptoms and signs of acromegaly within three years prior to hospitalization, including enlargement of the hands, feet, nose and jaw, thicker skin, deepening of the voice, and myocardial hypertrophy, etc. (  Figure 1A  ). T1 and T2 signals, and showed significantly inhomogeneous enhancement. The MRI also showed an invasive behavior with mass effect on the optic chiasm, left cavernous sinus involvement, and internal carotid artery encased (  Figure 1B  ). However, due to the hard texture of the tumor encircling the left internal carotid artery, some of the tumor body remained (  Figure 1C  ), and the postoperative serum GH level still did not drop to normal level which was about 7ng/mL (0.06-5.00ng/mL). Time line of the clinical course was shown in  Figure S1 .
(A) Hematoxylin-eosin HE revealed cap-like gangliocytoma cells surrounding the adenoma cells. (B) Immunohistochemistry showed strongly positive for growth hormone. (C) Immunohistochemistry showed strongly positive for synapsin. (D) Immunohistochemistry showed scattered positive for Prolactin (PRL). (E) Immunofluorescent staining show strong positive for Pituitary Transcript Factor 1(Pit-1). (F) The ultrastructure observed by Electron microscope.
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Figure 2A  ). Further immunohistochemistry profile showed strongly positive for GH and Synapsin, scattered positive for Prolactin(PRL) and Ck8/18, and about 5% monoclonal antibody (MIB)-1 proliferative index positivity (  Figures 2B   -D  ). Immunofluorescent staining show strong positive for Pituitary transcript factor 1(Pit-1) (  Figure 2E  ), which further proved its Pit-1 lineage origin. Further, the ultrastructure observed by electron microscope showed that tumor cells were densely distributed, with nuclei of different sizes, intracytoplasmic fibrous vesicles, round secretory granules within the fibrous vesicles, and scattered capillaries in the interstitium (  Figure 2F  ).
(A) Copy number variation (CNV) profiles of the two specimens: first surgery(scPA1) and second surgery(scPA2), show gain in chromosome 4 (both), and chromosome 11 deletion in scPA1 only, indicated the possible existence of temporal and spatial heterogeneity within the tumor. (B) Principal component analysis (PCA) showed that the cells were almost clustered together, and only little part of cells (7/209) had cell cycle heterogeneity (PC2 represented cell cycle related genes). (C) GO enrichment analysis of the differentially expressed genes of this case, the top 30 biological processes (BP) term were listed.
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Transcription sequencing (STRT-seq) technique was applied and the single-cell RNA sequencing (scRNA-seq) data was obtained through Illumina sequencing. Then, information on chromosome copy number variation (CNV) was obtained by the InferCNV method based on scRNA-seq data, and a comparison of the CNV profiles of the two specimens revealed that two specimens had gain in chromosome 4, while chromosome 11 deletion was only detected in the specimen obtained from the first surgery, thus suggesting the possible existence of temporal and spatial heterogeneity within the tumor (  Figure 3A  ). Then, all the 209 cells were analyzed by principal component analysis (PCA), and the results showed that the cells were almost clustered together, and only little part of cells (7/209) had cell cycle heterogeneity, which indicated that the heterogeneity within the tumor was small and about 3% of cells was in the process of mitosis (  Figure 3B  ). At last, the differentially expressed genes of this case were analyzed and the biological processes (BP) corresponding to them were clarified by Gene Ontology (GO) enrichment analysis, in which the biological processes that were more involved were "Cytoskeleton Organization", "anatomical structure development", and "cellular component assembly involved in morphogenesis"(  Figure 3C  ).