Table 1.

Clinical and morphologic features of the enrolled patients.

Pat. no.	Clinical features				Morphologic features
	Sex/Agea	MPN subtypeb	Gene carrying the driver mutation	Additional somatic mutationsc/High-molecular risk statusd	Light microscopy		Immunofluorescence microscopy
					Platelet anisocytosis	RBC anisopoikilocytosis	Platelet alpha granule defecte	Platelet dense granule defectf	Platelet cytoskeleton defectg	Platelet surface receptor defecth	RBC NMMIIA aggregatesi
1	M/73	PV	JAK2	No	n.a.	n.a.	Yes	Yes	No	No	No
2	F/53	MF	CALR	No	n.a.	n.a.	Yes	Yes	No	No	Yes
3	M/77	PV	JAK2	Yes/No	n.a.	n.a.	No	Yes	No	No	Yes
4	M/81	PV	JAK2	Yes/No	n.a.	n.a.	Yes	Yes	Yes	No	Yes
5	F/47	ET	CALR	Yes/No	n.a.	n.a.	Yes	Yes	Yes	No	Yes
6	F/66	ET	JAK2	No	n.a.	n.a.	Yes	Yes	Yes	No	Yes
7	M/55	ET	JAK2	No	n.a.	n.a.	Yes	Yes	No	No	Yes
8	M/36	MF	CALR	No	n.a.	n.a.	Yes	Yes	No	Yes	Yes
9	F/79	PV	JAK2	Yes/No	n.a.	n.a.	Yes	No	Yes	No	No
10	F/71	MF	JAK2	Yes/Yes	n.a.	n.a.	Yes	No	Yes	No	Yes
11	F/54	MPN-U	JAK2	No	n.a.	n.a.	No	No	No	No	Yes
12	M/44	ET	JAK2	No	n.a.	n.a.	No	Yes	Yes	No	Yes
13	F/79	MF	CALR	Yes/No	n.a.	n.a.	Yes	Yes	No	No	Yes
14	F/39	ET	CALR	No	n.a.	n.a.	No	No	No	No	No
15	F/61	PV	JAK2	Yes/No	n.a.	n.a.	Yes	No	No	No	No
16	M/59	MF	JAK2	No	n.a.	n.a.	Yes	Yes	Yes	No	Yes
17	M/80	PV	JAK2	Yes/No	n.a.	n.a.	Yes	No	Yes	No	Yes
18	M/63	ET	JAK2	Yes/No	n.a.	n.a.	Yes	No	No	Yes	Yes
19	M/40	PV	JAK2	No	n.a.	n.a.	No	No	No	No	Yes
20	F/63	ET	JAK2	Yes/No	n.a.	n.a.	Yes	Yes	No	No	Yes
21	F/69	MF	JAK2	No	n.a.	n.a.	Yes	No	No	No	Yes
22	M/67	MF	CALR	Yes/Yes	n.a.	n.a.	No	No	No	No	No
23	F/62	MF	CALR	No	n.a.	n.a.	No	Yes	No	No	Yes
24	M/78	MF	CALR	No	n.a.	n.a.	No	Yes	No	No	Yes
25	F/57	ET	CALR	Yes/No	n.a.	n.a.	No	No	No	Yes	Yes
26	M/58	PV	JAK2	Yes/No	n.a.	n.a.	No	No	No	No	Yes
27	F/43	MF	JAK2	No	n.a.	n.a.	No	Yes	No	No	No
28	M/70	PV	JAK2	Yes/No	n.a.	n.a.	Yes	No	No	No	Yes
29	M/79	MF	CALR	No	n.a.	n.a.	No	No	No	No	No
30	M/69	MF	JAK2	Yes/Yes	n.a.	n.a.	No	Yes	No	No	Yes
31	M/69	ET	JAK2	Yes/No	n.a.	n.a.	No	No	Yes	No	No
32	F/67	MF	JAK2	Yes/Yes	n.a.	n.a.	Yes	No	No	No	Yes
33	M/62	MF	JAK2	No	n.a.	n.a.	Yes	No	No	No	Yes
34	F/69	ET	JAK2	Yes/No	n.a.	n.a.	No	No	No	No	No
35	M/55	MF	CALR	n.a./n.a.	n.a.	n.a.	No	No	Yes	No	Yes
36	M/52	MF	CALR	No	n.a.	n.a.	No	No	No	No	Yes
37	F/68	MPN-U	TN	Yes/-	n.a.	n.a.	No	No	No	No	No
38	F/27	ET	JAK2	No	n.a.	n.a.	No	No	No	Yes	No
39	F/34	ET	JAK2	No	n.a.	n.a.	No	Yes	No	No	No
40	F/52	PV	JAK2	Yes/No	Yes	No	Yes	Yes	No	No	Yes
41	F/48	MF	JAK2	No	Yes	No	Yes	Yes	Yes	No	Yes
42	M/68	MF	JAK2	Yes/Yes	Yes	Yes	Yes	Yes	No	No	Yes
43	F/51	PV	JAK2	No	No	No	Yes	No	No	No	No
44	F/22	MF	JAK2	No	Yes	No	Yes	No	No	No	Yes
45	M/58	MF	JAK2/MPL	Yes/Yes	Yes	Yes	Yes	Yes	Yes	No	Yes
46	M/82	MF	JAK2	Yes/No	Yes	No	Yes	No	No	No	Yes
47	M/71	PV	JAK2	No	n.a.	n.a.	Yes	Yes	No	No	No
48	M/67	MF	MPL	Yes/No	Yes	Yes	Yes	No	No	No	Yes
49	M/69	PV	JAK2	Yes/No	Yes	No	Yes	Yes	No	No	No
50	F/64	PV	JAK2	No	Yes	No	No	Yes	No	No	No
51	F/52	MF	JAK2	No	Yes	No	Yes	No	No	No	No
52	F/55	MF	JAK2	Yes/No	No	Yes	Yes	No	No	No	Yes
53	M/81	PV	JAK2	No	n.a.	n.a.	Yes	No	No	No	Yes
54	M/75	MF	JAK2	No	Yes	No	No	No	No	No	Yes
55	F/68	MF	JAK2	Yes/No	No	No	No	No	No	No	No
56	M/48	ET	JAK2	Yes/No	Yes	No	Yes	No	No	No	Yes
57	M/80	MF	JAK2	No	Yes	Yes	Yes	Yes	No	No	Yes
58	F/62	MF	JAK2	Yes/Yes	Yes	Yes	Yes	No	No	No	Yes
59	F/68	MF	CALR	No	Yes	Yes	No	No	No	No	Yes
60	M/64	MF	CALR	Yes/Yes	Yes	Yes	No	No	No	No	Yes
61	F/62	PV	JAK2	No	Yes	No	No	No	No	No	No
62	F/59	ET	JAK2	No	Yes	No	No	No	No	No	No
63	F/39	MF	MPL	No	Yes	Yes	Yes	No	No	No	Yes
64	F/74	ET	JAK2	Yes/No	Yes	No	No	No	No	No	Yes
65	M/77	MPN-U	JAK2	Yes/-	Yes	No	Yes	Yes	No	No	Yes
66	F/53	PV	JAK2	Yes/No	Yes	No	No	No	No	No	Yes
67	M/77	ET	MPL	Yes/No	Yes	Yes	Yes	Yes	No	No	Yes
68	F/63	MF	JAK2	No	Yes	Yes	Yes	No	No	No	Yes
69	F/73	PV	JAK2	Yes/No	No	No	No	Yes	No	No	Yes
70	M/51	MF	CALR	Yes/No	Yes	Yes	Yes	No	No	No	Yes
71	M/64	MF	JAK2	Yes/No	Yes	Yes	Yes	No	No	No	Yes
72	F/67	MF	TN	Yes/Yes	Yes	Yes	No	Yes	No	No	Yes
73	M/47	MF	MPL	Yes/Yes	Yes	No	Yes	Yes	No	No	No
74	F/63	PV	JAK2	No	No	No	Yes	Yes	No	No	No
75	F/33	PV	JAK2	No	Yes	No	No	No	No	No	No
76	M/46	PV	JAK2	Yes/No	No	Yes	No	No	No	No	Yes
77	F/59	ET	JAK2	No	Yes	Yes	Yes	No	No	No	No
78	F/75	MF	JAK2	Yes/No	Yes	No	Yes	No	Yes	No	Yes
79	M/54	MF	MPL	No	No	No	No	No	No	No	Yes
80	F/78	MF	CALR	Yes/No	Yes	No	Yes	No	Yes	No	Yes
81	F/80	ET	MPL	No	Yes	No	Yes	No	Yes	No	Yes
82	F/31	PV	JAK2	No	No	No	No	No	Yes	No	Yes
83	M/60	PV	JAK2	Yes/No	Yes	No	No	No	Yes	No	Yes
84	M/44	ET	CALR	Yes/No	No	No	Yes	No	No	No	No
85	F/43	MF	JAK2	No	Yes	Yes	Yes	Yes	Yes	No	Yes
86	F/45	PV	JAK2	Yes/No	No	No	Yes	Yes	No	No	No
87	M/65	MF	JAK2	Yes/No	Yes	Yes	No	Yes	No	No	Yes
88	F/39	MF	JAK2	Yes/No	No	No	No	No	No	No	Yes
89	F/64	MPN-U	JAK2	Yes/-	No	No	Yes	No	No	No	Yes
90	F/39	ET	CALR	No	Yes	Yes	No	Yes	No	No	Yes
91	F/84	PV	JAK2	Yes/No	Yes	Yes	Yes	No	No	No	Yes
92	M/54	MF	CALR	Yes/Yes	Yes	Yes	No	No	No	No	Yes
93	F/40	ET	JAK2	No	Yes	Yes	Yes	Yes	Yes	No	Yes
94	F/82	MPN-U	JAK2	Yes/-	Yes	Yes	Yes	Yes	No	No	Yes
95	F/58	PV	JAK2	No	Yes	No	Yes	No	No	No	Yes
96	M/60	PV	JAK2	No	Yes	Yes	Yes	Yes	No	No	Yes
97	M/58	ET	JAK2	No	Yes	No	No	No	No	No	Yes
98	M/79	ET	MPL	Yes/No	Yes	Yes	Yes	Yes	No	No	Yes
99	F/76	MPN-U	TN	Yes/-	No	No	No	No	No	No	No
100	M/31	MF	TN	No	No	No	Yes	Yes	Yes	No	Yes
101	F/71	MF	CALR	Yes/Yes	Yes	No	No	No	No	No	No
102	M/71	ET	JAK2	n.a./n.a.	Yes	Yes	Yes	Yes	Yes	No	Yes
103	F/60	PV	JAK2	No	Yes	No	No	No	No	No	No
104	F/78	MF	JAK2	Yes/Yes	Yes	Yes	Yes	No	No	No	Yes
105	F/80	MF	JAK2	Yes/No	Yes	Yes	Yes	Yes	No	No	Yes
106	M/73	PV	JAK2	Yes/No	Yes	Yes	Yes	Yes	No	No	Yes
107	F/74	MF	CALR	No	Yes	No	Yes	Yes	No	No	Yes
108	F/43	MPN-U	JAK2	Yes/-	Yes	No	Yes	Yes	No	No	Yes
109	M/68	MF	MPL	Yes/Yes	Yes	Yes	Yes	Yes	No	No	Yes
110	M/80	PV	JAK2	Yes/No	No	Yes	Yes	Yes	No	No	Yes
111	M/45	PV	JAK2	No	Yes	No	Yes	Yes	No	No	Yes
112	M/79	MF	JAK2	Yes/No	n.a.	n.a.	Yes	Yes	No	No	Yes
113	M/74	MF	JAK2	Yes/No	Yes	No	No	Yes	No	No	No
114	F/39	ET	JAK2	No	n.a.	n.a.	No	Yes	No	No	No
115	F/80	MF	JAK2	No	Yes	Yes	Yes	Yes	No	No	Yes
116	F/67	PV	JAK2	No	No	No	Yes	Yes	No	No	Yes
117	F/68	ET	JAK2	Yes/Yes	No	Yes	Yes	Yes	No	No	Yes
118	M/50	ET	JAK2	Yes/No	Yes	Yes	No	Yes	No	No	Yes
119	F/71	MF	JAK2	Yes/Yes	No	Yes	Yes	No	No	No	Yes
120	M/75	MF	JAK2	No	Yes	Yes	Yes	No	Yes	No	Yes
121	F/62	MF	CALR	Yes/No	Yes	Yes	No	No	No	No	Yes
122	M/67	MF	JAK2	Yes/Yes	Yes	No	Yes	Yes	No	No	Yes
123	M/55	MF	CALR	No	Yes	Yes	Yes	No	Yes	No	Yes
124	F/45	PV	JAK2	Yes/No	Yes	No	Yes	Yes	No	No	Yes
125	F/61	MF	JAK2	Yes/No	Yes	No	No	No	No	No	No
126	M/28	MF	CALR	No	Yes	No	No	No	No	No	No
127	F/79	PV	JAK2	No	Yes	No	Yes	Yes	No	No	Yes
128	F/48	ET	JAK2	Yes/No	Yes	No	Yes	No	No	No	No
129	M/87	MPN-U	JAK2	Yes/-	Yes	No	Yes	No	No	No	Yes
130	F/60	MF	JAK2	Yes/No	Yes	Yes	Yes	Yes	Yes	Yes	Yes
131	F/47	ET	JAK2	Yes/No	Yes	No	No	Yes	No	No	No
132	F/61	ET	CALR	Yes/No	Yes	Yes	Yes	No	No	No	No
133	M/62	ET	CALR	Yes/No	Yes	No	No	No	No	No	Yes
134	F/44	PV	JAK2	Yes/No	No	No	No	No	No	No	No
135	M/70	PV	JAK2	n.a./n.a.	No	Yes	Yes	Yes	No	No	Yes

Pat. patient, No. number, F female, M male, MPN myeloproliferative neoplasm, PV polycythemia vera, ET essential thrombocythemia, MF primary or secondary myelofibrosis, MPN-U unclassifiable MPN, TN triple-negative, i.e., absence of mutations hitting either JAK2 or CALR or MPL; n.a. not available, RBC red blood cell, NMMIIA non-muscular myosin IIA.

^aAt time of investigation.

^bAccording to the WHO 2016 classification [13].

^c≥1 additional somatic mutation detected by NGS analysis of 33 genes (ASXL1, BCOR, CBL, CEBPA, CUX1, DNMT3A, EZH2, GATA2, GNAS, GNB1, IDH1, IDH2, NF1, PHF6, PHIP, PPM1D, PRPF8, PTPN11, RAD21, RAS, RB1, RUNX1, SETBP1, SF3B1, SH2B3, SMC1A, SMC4, SRSF2, STAG2, TET2, TP53, U2AF1, ZRSR2) with a variant allele frequency ≥ 2%.

^d≥1 high-molecular risk mutation according to the prognostic panels of PV, ET and PMF [11, 12] - where applicable.

^eDefined as reduced expression of at least two out of the three investigated markers of alpha granules (von Willebrand factor, P-selectin, thrombospondin 1) compared to control, as reported [8].

^fDefined as reduced or altered expression of at least two out of the three investigated markers of lysosomes and dense granules (LAMP-1, LAMP-2, CD63) compared to control, as reported [7].

^gDefined as altered expression of at least three out of the four investigated cytoskeletal proteins (filamin A, NMMIIA, α-tubulin, β1-tubulin) compared to control, as reported [7].

^hDefined as reduced expression of the surface glycoprotein Ib/IX or IIb/IIIa compared to control, as reported [7];

ⁱDetection of NMMIIA aggregates in RBC, as reported [8]. Pat. = patient; No. = number; F = female; M = male; MPN = myeloproliferative neoplasm; PV = polycythemia vera; ET = essential thrombocythemia; MF = primary or secondary myelofibrosis; MPN-U = unclassifiable MPN; TN = triple-negative, i.e., absence of mutations hitting either JAK2 or CALR or MPL; n.a. = not available; RBC = red blood cell; NMMIIA = non-muscular myosin IIA.