JC THYROID B-7 H-3 and ICAM-1 are .pptx

Thyroid malignancy

1. B7-H3 AND ICAM-1 ARE POTENTIALLY THERAPEUTIC TARGETS FOR THYROID CARCINOMA PRESENTED BY : Dr Farheen khan MODERATED BY : Dr Sudarshana mam

2. ANATOMY - THYROID GLAND Butterfly shaped endocrine gland composd of 2 lateral lobes connected by a thin band of tissue called as isthmus. Location - In neck, in front of the upper part of trachea below the level of the cricoid cartilage C5 -T1 vertebrae. Normal weight - 15- 25 grams

3. BLOOD SUPPLY- Superior thyroid artery Inferior thyroid artey VENOUS SUPPLY- Superior thyroid veins Middle tyroid veins Inferior thyroid veins LYMPHATIC DRAINAGE Pericapsular Internal jugular chain Pretracheal, paratracheal, and prelaryngeal ( Delphian node) Recurrent laryngeal nerve chain; retropharyngeal and retroesophageal .

4. NORMAL HISTOLOGY 2 types of cells - Follicular cells Parafollicular cells or C cells THYROID FOLLICLE - is the FUNCTIONAL UNIT of thyroid gland. It consists of a single layer of COLUMNAR to CUBOIDAL epithelial cells resting on a basement membrane. It encloses amorphous material called as COLLOID These are surrounded by rich capillary network. About 20-40 follicles are demarcated by connective tissue septa to form a lobule. Cytoplasm contains extensive endoplasmic reticulum.

5. Follicular cells - synthesize and secretes iodine containing hormones T3 and T4 Colloid - contains mainly Thyroglobulin(Tg) that stores the thyroid hormones before secretion. Parafollicular cells - synthesize and secrete the hormone calcitonin

6. Functions of thyroid hormones Bone growth Maturation of central nervous system Beta - adrenergic actions - increased c.o ,heart rate,and contractility of myocardium Increased basal metabolic rate Increased glycogenolysis,gluconeogenesis and lipolysis

7. THYROID TUMORS Clinically apparent thyroid neoplasms - Primary and Epithelial Traditionally - Adenomas and carcinomas From a histogenetic/differentiation standpoint - divided into 3 major categories - Tumors exhibiting follicular cell differentiation Tumors exhibiting C- cell differentiation Tumors exhibiting follicular and C- cell differentiation.

8. PAPILLARY CARCINOMA OF THYROID I t i s t he m o s t c o mm o n t y p e o f t h yr o i d c a r c i no m a and M o s t c o m m on e nd oc r i n e malignancy comprising of 75-85% of cases. Papillary thyroid carcinoma (PTC) is a malignant epithelial tumour showing evidence of follicular cell differentiation and a set of distinctive nuclear features. Papillary carcinoma is defined by the presence of a distinctive set of alterations of nuclear m o r p ho l o g y , although t he p r e s e n c e of e i t h e r papillae or invasion of the surrounding thyroid parenchyma is generally required for the diagnosis.

9. CLINICAL FEATURES Nodule or mass in the neck. Dysphagia or hoarsness (due to compression). Weight loss and difficulty in breathing Females are more affected than males. It can present in any age group, mainly 40–50 years . It accounts for more than 90% of thyroid malignancies in children. In approximately 5% of cases, there is a history of irradiation exposure to the neck. There is increase in the incidence of papillary carcinoma in patients of hashimoto thyroiditis.

10. ETIOLOGY Conventional PTC have two defining genetic abnormalities: translocations that result in gene fusions of RET OR NTRK (neurotrophic tyrosine receptor kinase) and point mutations in BRAF. The RET gene is located on chromosome 10q11, and encodes an RTK (receptor tyrosine kinase) that is normally not expressed in thyroid follicular cells. PTC rearrangements involving RET create fusion genes t hat encode constitutively active forms of RET tyrosine kinase. There are more than 15 fusion partners of RET , and two designated PTC 1 and PTC 2 —are most commonly observed in sporadic papillary cancers. These rearrangements also produce constitutively active NTRK 1 fusion proteins.

11. CONTD… In BRAF gene , most common change is valine-to-glutamate change in codon 600 ( brafv600e ). Braf encodes a serine/threonine kinase that lies downstream of receptor tyrosine kinases in growth factor signaling pathways. The presence of BRAF and RET mutations in C onventional PTC is associated with reduced expression of thyroid differentiation markers (such as thyroglobulin and thyroid peroxidase ) and may be associated with a higher risk of extrathyroidal extension and recurrence.

12. GROSS Grossly, PTC typically presents as an invasive neoplasm with poorly defined margins, a firm consistency, and a granular white cut surface. Calcifications may be apparent. The size is widely variable, with a mean diameter of 2-3 cm. Sometimes marked cystic changes are seen.

13. MICROSCOPY The two cardinal morphological features of conventional (classic) PTC are the papillae and the nuclear changes. The papillae are composed of a central fibrovascular stalk covered by a neoplastic epithelial lining. The papillae may be long, straight, or arborizing; arranged in a parallel, regimented fashion; short and stubby; or tightly packed. The stalk is usually made up of loose connective tissue and variously sized thin-walled vessels

14. CONTD…. The nuclei of the cells of PTC display a characteristic set of abnormalities, which can be grouped into three categories: (1) changes in size and shape, (2) irregularities of the membrane, and (3) chromatin characteristics Changes in nuclear size and shape include nuclear enlargement and often overlapping, as well as nuclear elongation. .

15. CONTD…. The nuclei of PTC are typically characterized by marked remodelling of the nuclear envelope, with highly irregular nuclear contours and nuclear pseudoinclusions or prominent longitudinal grooves . Nuclei is centrally located with finely dispersed chromatin and an optically clear or empty cytoplasm , giving rise to the designation ground-glass or orphan annie eye nuclei. Small, concentrically calcified structures termed psammoma bodies are often present, usually within the cores of papillae. Metastases to adjacent cervical lymph nodes occur in up to one- half of cases.

16. HISTOCHEMICAL AND IMMUNOHISTOCHEMICAL FEATURES Thyro-globulin and TTF-1 are very specific markers. PAX-8 can be used to differentiate papillary carcinoma (positive for both ttf-1 and pax8) from lung adenocarcinoma and other tumors that are positive for ttf-1 but negative for pax8.

17. Molecular Genetic Features Activation of the mitogen-activated protein kinase ( mapk ) pathway is the defining molecular genetic feature of papillary carcinoma. This pathway regulates important cellular functions (E.G., Proliferation, differentiation, and survival) and is frequently altered in human cancers.

18. VARIANTS OF PAPILLARY CARCINOMA OF THYROID A- Encapsulated variant - A papillary carcinoma totally surrounded by a Fibrous capsule, which may be intact or only focally infiltrated by tumour growth. The encapsulated variant accounts for about 10% of all cases of PTC. Characterized microscopically by a pale, vacuolated colloid. The papillary areas are largely limited to the area facing the cystic cavity. Encapsulated PTC has an excellent prognosis. Regional nodal metastases may be present. .

19. B- Oncocytic (Oxyphilic, Warthin-Like) Variant- In this variant, the nuclear features remain those of papillary carcinoma, but the cytoplasm is abundant and has a granular oxyphilic quality. The prognosis of Warthin-like papillary carcinomas is generally very good.

20. C- Follicular variant of PTC- This variant has an exclusively or almost exclusively follicular growth pattern. Most importantly, the nuclei of the cells lining the follicles have features of conventional PTC. The colloid within the lumina of the neoplastic follicles often has a strong and homogeneous eosinophilic quality. Also called as lindsay tumor.

21. D. Diffuse sclerosing variant This uncommon variant occurs more commonly in women, most frequently in the second or third decade of life. Elevated serum antithyroglobulin and antimicrosomal antibodies may mimic Hashimoto thyroiditis and a hard so-called woody gland may mimic Riedel thyroiditis. Histologically, there is diffuse involvement of a single lobe or the entire gland, characterized by dense sclerosis, numerous psammoma bodies, and background changes of chronic lymphocytic thyroiditis. Tumour nests appear solid, with associated squamous metaplasia, including squamous morules . The tumour cells have a propensity to invade intrathyroidal lymphatic spaces and a tendency to show extrathyroidal extension.

22. E. Tall cell variant This variant is composed of cells that are two to three times as tall as they are wide, and that show abundant eosinophilic ( oncocytic -like) cytoplasm. Typical nuclear features of papillary carcinoma are present, and nuclear pseudoinclusions are usually easily found. Tall cells must account for > 30% of all tumour cells for the diagnosis of the tall cell variant . The tall cell variant of PTC occurs at an older patient age and is considered an aggressive variant.

23. MEDULLARY CARCINOMA A distinctive type of malignant thyroid neoplasm composed of C (parafollicular) cells Prior synonyms - solid carcinoma with amyloid stroma, hyaline carcinoma, and C-cell carcinoma Medullary microcarcinoma - small tumors (<1 cm) with a low risk of recurrence and metastasis.

24. GROSS Grossly, the typical tumor is solid, firm, and nonencapsulated but relatively well circumscribed and has a gray to yellowish cut surface. A continuous fibrous capsule envelops an occasional tumor. Most tumors are located in the midportion or upper half of the gland, corresponding to a greater concentration of C cells in these regions

25. MICROSCOPY Microscopically, the classic medullary carcinoma cytoarchitecture is represented by a solid proliferation of round to polygonal cells of granular amphophilic cytoplasm and medium-sized nucleus, separated by a highly vascular stroma, hyalinized collagen, and amyloid The pattern of growth can be carcinoid-like,paraganglioma-like, trabecular, glandular (tubular and follicular),or pseudopapillary The stroma may be scanty, hemorrhagic, ossified, or edematous. The amyloid deposition may be widespread, limited to small psammomatoid concretions, or altogether absent The tumor cells may be plasmacytoid (because of nuclear peripheralization), spindle shaped, oncocytic, squamoid, or squamous The oncocytic variety is particularly treacherous because of its uncanny resemblance to Hürthle cell (oncocytic)

26. CYTOLOGIC FEATURES In FNA preparation - It is characterized by eccentric nuclei, salt and pepper (neuroendocrine type) chromatin, inconspicuous nucleoli, binucleated and multinucleated cells, ill-defined cell borders, and a clean background.Amyloid material can be identified in some cases

27. ELECTRON MICROSCOPY cytoplasmic dense-core secretory granules are invariably present. A variability in their sizes has been detected, suggesting the existence of multiple endocrine cell types in this tumor. These granules are usually argyrophilic with the Grimelius stain, especially if the tissue has been fixed in Bouin fluid.

28. IHC Keratin Thyroid markers - TTF-1 Parathyroid markers -chromogranin A, B,and C synaptophysin;CD56;opioid peptides Calcitonin - However, occasional medullary carcinomas display either focal or complete loss of calcitonin expression (a diagnostic pitfall). They are also consistently positive for CEA and negative for thyroglobulin small peptide products : somatostatin, adrenocorticotropic hormone (ACTH), calcitonin gene–related peptide, serotonin, melanocyte-stimulating hormone (MSH), prostaglandins, bombesin, gastrin-releasing peptide, substance P, L-dopa decarboxylase, hista_x0002_minase, glucagon, insulin, hCG, the polysialic acid of the neural cell adhesion molecule, galectin-3, hepatocyte growth factor and its receptor, matrix metalloproteinases, prohormone convertases, and progesterone receptors

29. ANAPLASTIC (Undifferentiated) CARCINOMA typically presents in elderly patients as a rapidly growing firm mass, fixed to the surrounding structures, associated with hoarseness, dysphagia, and dyspnea. Extrathyroidal extension is encountered at the time of initial presentation in most of the cases

30. GROSS Grossly, a highly necrotic and hemorrhagic solid tumor mass is seen replacing large portions of the organ

31. MICROSCOPY The first is undifferentiated in the sense that it does not make follicles, papillae, or even trabeculae or nests, but the tumor still retains an unmistakable epithelial appearance on morphologic and immunohistochemical grounds. This pattern, which is referred to as squamoid , may blend with clear-cut foci of keratinization. The second category is composed of two patterns which are often seen together and which can be grouped under the qualifier of sarcomatoid: spindle cell and giant cell One variation on the theme of the spindle cell form of undifferentiated carcinoma is the paucicellular variant, which mimics Riedel thyroiditis because of the extreme degree of fibrosis and hyalinization, and which is recognized because of the scattered atypia, areas of necrosis, vascular invasion, vascular permeation, and positivity for epithelial markers The rare rhabdoid variant may be considered a variation on the theme of the giant cell form of undifferentiated carcinoma.

32. IHC Immunohistochemically, the most useful marker for confirming the epithelial nature of the tumor is keratin (50-100%cases) Vimentin is consistently present in the neoplastic spindle cells focal EMA and CEA positivity may be found ( in squamoid type) TTF1 and TTF-2 (FoxE1) are positive in minority of anaplastic carcinomas PAX8 P53(diffusely positive) Has very high proliferative activity (Ki-67/MIB1 labeling index ~35%)

33. INTRODUCTION TO JOURNAL Although most differentiated thyroid carcinoma has a clinically favorable prognosis, some of specifc types of thyroid cancer (such as anaplastic thyroid carcinoma and advanced papillary thyroid carcinoma) show fatal outcomes and require novel treatments. Immunotherapy is a promising avenue for the treatment of advanced thyroid carcinoma. B7-H3 (B7 homolog 3 protein) and ICAM-1 (intercellular adhesion molecule 1), as two important immune checkpoints (ICPs), is becoming hopeful target spots for immunotherapy.

34. A growing amount of evidence has suggested that B7-H3 and ICAM-1 are upregulated in papillary thyroid carcinoma. In the present study, we explored the expression level of B7-H3 and ICAM-1 in diferent types of thyroid carcinoma. In the groups of the TCGA cohort, both B7-H3 and ICAM-1 mRNA were highly expressed in thyroid carcinoma.

35. Thyroid cancer is the most frequent type of endocrine system malignancy Categorised into 3 broad histological categories : 1) differentiated thyroid cancer -originate from thyroid follicular epithelial cells (papillary, oncocytic and follicular thyroid cancer) 2) Anaplastic thyroid cancer(ATC) 3) Medullary thyroid cancer (MTC)- originate from the parafollicullar neuroendocrine cells of the thyroid Most prevelant subtype - Papillary thyroid cancer(PTC)

36. PTC presents as a slow progressing tumor and carries the most favourable overall prognosis. Small proportions of thyroid cancers such as poorly differentiated PTC(PDPTC),MTC and ATC exhibit high aggressiveness and even lethality Conventional treatments for thyroid cancer include surgical intervention,radioactive iodine(RAI)therapy, and TSH suppression therapy. Recently targeted therapies and antiangiogenic multikinase inhibitors are approved for treatment of thyroid cancers. However,there is a paucity of effective treatment option available for patients with advanced thyroid cancer.

37. Previous studies have postulated that the distinct prognosis is a result of intricate interactions between tumor cells and their microenvironment. Indeed, immuno therapy utilizing immune checkpoint inhibitors targeting PD1, PD-L1, and CTLA4 has emerged as a well-established option for cancer treatment in recent decades. Although immunotherapy cannot currently replace the standard of care, immune-based approaches remain a promising avenue for treatment. In this study, we analyzed the expression level of two immune checkpoints (ICPs) B7H3 (B7 homolog 3 protein) and ICAM-1 (intercellular adhesion molecule 1) as well as their correlation between clinic-pathological features in thyroid cancer. Moreover, we initially evaluate the correlations between the levels of B7-H3 and ICAM1 in thyroid cancer.

38. MATERIALS AND METHODS Analysers of the UALCAN databases and GSE213647 In the present study, the UALCAN website is used to explore the expression levels of B7-H3 and ICAM-1 in thyroid cancer and normal thyroid tissues. Additionally, the correlation between B7-H3 and ICAM-1 in thyroid cancer is evaluated in GSE213647. STUDY TYPE- Cohort

39. Tissue samples collection Tissue arrays (HTyCan060PT01) containing 28 patients who underwent surgical resection for thyroid cancer were obtained from Shanghai Outdo Biotech. The study design was approved by the Institutional Review Board of Huzhou central Hospital/Shanghai QBC Ethics Committee. All pathologic specimens were independently certifed by two pathologists. Stage classifcation was assessed according to the TNM classification. The clinicopathologic characteristics of the patients are summarized in the given table 1.

40. Patient characteristics A total of 28 patients with thyroid cancer were enrolled,including 18 PTCs, 6 MTCs, 2 FTCs, 2 ATCs (among them one is SCC). Among them 9 were men and 19 were women. The mean age at diagnosis was 56 years. 71.4% (20/28) cases had well diferentiated tumor and 35.7% (10/28) patients had small tumor size (max diameter<2 cm). Among these patients, 3 patients showed signs of lymph node metastasis (N1) and no patients showed distant metastasis.

41. Immunohistochemical (IHC) staining of B7-H3 and ICAM-1 in thyroid cancer IHC studies were conducted in samples of the 28 thyroid cancer cases. Both B7-H3 (CD276) and ICAM-1(CD54) staining were mainly localized to the cytoplasmic /membranous of tumor cells The representative regions of IHC are shown in Figs. 2 and 3. Via IHC, B7-H3 (CD276) protein expression was found in 28 of 28 thyroid cancers (100%) and had a signifcantly higher expression of B7-H3 (CD276) expression in tumor tissues than in normal tissues (Fig.  2B). Among the positive cases, MTCs and FTCs exhibited relatively weaker expression of B7-H3 (CD276).

42. While, PTCs and ATCs showed frequently moderate to strong expression of B7-H3 (CD276). Moreover, compared with normal tissues, ICAM-1(CD54) was also increased in thyroid cancer tissues (Fig.  3B). The results showed that ICAM-1 (CD54) protein expression was detected in 27 thyroid cancer tissues, including low expression levels in 9 samples (33.3%) and high expression levels in 18 samples (66.7%). Overall, these data suggest that the expression of B7-H3 (CD276) and ICAM-1 (CD54) were up-regulated in thyroid cancer tissues (Fig. 4A,B). We also found that the patients with 61-80y had lower protein expression of both B7-H3 (CD276) and ICAM-1 (CD54). However, the lack of statistical difference between the two groups (41-60y and 61-80y) might be due to the small sample size and semi-quanti_x0002_tatively score (p=0.079 and p=0.567). However, there was no signifcant association between B7-H3 (CD276) or ICAM-1 (CD54) expression and patient’s gender (Fig. 4C).

43. Evaluation of IHC staining - A total of 56 TMA points were retained for further analysis. To quantify B7-H3 and ICAM-1 protein expression, all stained points were separately evaluated by two senior pathologists. Both the staining extensity and intensity were assessed in randomly selected five representative fields of vision. According to the percentage of positive tumor cells, B7-H3 and ICAM-1 protein expression were further stratifed into negative (<1%), low (1%~49%), and high (≥50%).

44. RESULT The expression level of B7-H3 and ICAM-1 mRNA in thyroid cancer - In the TCGA databases, there was a signifcant diference in B7-H3 (CD276) expression between normal thyroid tissue samples (n=59) and thyroid cancer tissue samples (n=505) (p<0.001, Fig.  1A), B7-H3 (CD276) is highly expressed in tumor tissues. Furthermore, the expression level of B7H3 (CD276) was signifcantly diferent in histology, stages, patient’s age and nodal metastasis status (Fig. 1B-E). Statistical analysis indicated that the patients with Stage2, 61-80y, Follicular thyroid papillary carcinoma and N0 had lower B7-H3 (CD276) mRNA expression. there was no signifcant association between B7-H3 (CD276) expression and patient’s gender (Fig. 1F).

45. Similar to B7-H3, the same result was also obtained in the analysis of ICAM-1(CD54) mRNA and thyroid cancer tissue samples (Fig. 1G-L). These data indicated that the level of B7-H3 (CD276) and ICAM-1(CD54) mRNA are up-regulated in thyroid cancer tissues.

46. CONCLUSION In this research, we are the first to show the co-expression of ICAM1 and B7H3 mRNA as well as their associations between clinico-pathological features in thyroid cancers. Furthermore, the correlation between ICAM1 and B7H3 were also evaluated. The signifcant relevance of B7-H3 staining score with ICAM-1 staining score was observed in TCGA database and our cohort, which might open avenues for the combination therapy in advanced thyroid cancer. If the interacting mechanism between ICAM1 and B7H3 can be full identifed we might open avenues for the combination therapy in advanced thyroid cancer (Fig. 5).

47. THANK YOU JEFF FOSKETT, american singer Died due to Anaplastic thyroid cancer on 11 dec 2023