Evaluation of LS301 Uptake in Tumors of Patients Undergoing Partial Mastectomy and Sentinel Lymph Node Biopsy for Breast Cancer
Phase: PHASE1
Conditions: Breast Cancer, Cancer of the Breast
Sponsor: Samuel Achilefu

S T U D Y P R O T O C O L A N D S T A TI S TI C A L A N A L Y SI S P L A N  
 
 
 
Offi ci al titl e:  E v al u ati o n of L S 3 0 1 U pt a k e i n T u m or s of 
P ati e nt s U n d er g oi n g P arti al M a st e ct o m y a n d S e nti n el L y m p h 
N o d e Bi o p s y f or Br e a st C a n c er  
 
N C T n u m b er:  N C T 0 2 8 0 7 5 9 7  
I R B A p pr o v e d d at e : 0 6 -0 6 - 20 2 3  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 of 5 4  
 
 
 
 
 
E v al u ati o n of L S 3 0 1 U pt a ke i n T u m ors of P atie nts U n der g oi n g P arti al M astect o m y a n d 
Se nti nel L y m p h N o de Bi o ps y f or Bre ast C a ncer  
 
 
P h ase 1 A: 
Was hi n gt o n U ni versit y Sc h o ol of Me dici ne 
De part me nt of S ur g er y  
6 6 0 S o ut h E ucli d A ve n ue, Ca m p us B o x 8 1 0 9 
St. L o uis, M O  6 3 1 1 0 
 
P h ase 1 B/ P h ase 2:  
U T S o ut h wester n Me dical Ce nter  
5 3 2 3 Harr y Hi n es Bl v d.  
Dallas, T X 7 5 3 9 0 
 
Pr ot oc ol #: 2 0 1 9 0 7 1 3 5  
Versi o n D ate: 0 5/ 1 5/ 2 0 2 3  
S T U - 2 0 2 2- 0 3 8 5 
 
  
P h ase 1 A: 
Pri nci p al I n vesti g at or:  
 
J ulie Mar ge nt haler , M. D. 
P h o ne:  ( 3 1 4) 3 6 2- 7 5 3 4 
E m ail:  j mar ge nt haler @ w ustl.e d u  
 
P h ase 1 B & P h ase 2 : 
Pri nci p al I n vesti g at or:  
 
De b ora h Farr, M. D. 
  
  
  
  
 
U T S o ut h wester n Me dical Ce nter  
  
  
  
  
 
De part me nt of S ur g er y  
  
  
  
  
  
5 3 2 3 Harr y Hi n es Bl v d., Dallas, T X 7 5 3 9 0 
  
  
  
  
 
P h o ne:  ( 2 1 4) 6 4 8- 5 8 7 0 
  
  
  
  
 
E m ail:  De b ora h.farr @ uts o ut h wester n.e d u  
 
P h ase 1 A: 
C o - Pri nci p al I n vesti g at or:   
Sa m uel Ac hilef u, P h D 
U T S o ut h wester n Me dical Ce nter  
  
  
  
  
 
De part me nt of Bi o me dical E n gi neeri n g 
5 3 2 3 Harr y Hi n es Bl v d., Dallas, T X 7 5 3 9 0 
P h o ne:  ( 3 1 4) 3 6 2- 8 5 9 9 
E m ail:  ac hilef u @ w ustl.e d u  
 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 of 5 4  
 
 
P h ase 1 B & P h ase 2: 
C o - Pri nci p al I n vesti g at ors or Le a d S u b-I n vesti g at or(s):   
 
  
  
  
  
 
Sa m uel Ac hilef u, P h D 
U T S o ut h wester n Me dical Ce nter  
  
  
  
  
  
De part me nt of Bi o me dical E n gi neeri n g 
5 3 2 3 Harr y Hi n es Bl v d., Dallas, T X 7 5 3 9 0 
                                                           P h o ne:  2 1 4- 6 4 5- 9 9 0 0 
                                                           E m ail:  Sa m uel. Ac hilef u @ U T S o ut h wester n.e d u  
  
 
P h ase 1 A: 
S u b -I n vesti g at or(s):    
  
  
 
S u ma n M o n dal, P h. D.  
  
  
Ra di ol o g y 
C hristi ne O’ Brie n, P h. D.  
  
  
Ra di ol o g y 
R ui Ta n g, P h. D.  
  
  
  
Ra di ol o g y 
Xia o X u, P h. D.  
  
  
  
Ra di ol o g y 
Ji n g qi n ( R os y) L u o, P h. D.  
  
  
Bi ostatistics 
Ti m ot h y E berlei n, M. D.  
  
  
S ur gical O nc ol o g y 
Willia m E. Gilla n ders, M. D.     
  
S ur gical O nc ol o g y 
Re becca L. Aft, M. D., P h. D.    
  
S ur gical O nc ol o g y 
Kat heri ne Gl o v er - C olli ns, M. D., P h. D.  
S ur gical O nc ol o g y 
De b ora h Veis ( N o v ac k), M. D., P h. D.  
Pat h ol o g y 
 
P h ase 1 B & P h ase 2 : 
S u b -I n vesti g at or(s):   
N/ A     
 
                                                               
 
Bi ost atistici a n:   
 
Mi nJae Lee, P h D  
  
  
  
 
U T S o ut h wester n Me dical Ce nter  
  
  
  
  
5 3 2 3 Harr y Hi n es Bl v d., Dallas, T X 7 5 3 9 0 
  
  
  
 
P h o ne:  ( 2 1 4) 6 4 8- 3 8 0 3 
  
  
  
 
F a x:  ( 2 1 4) 6 4 8- 3 9 3 4 
  
  
  
  
Mi nJae. Lee @ U T S o ut h wester n.e d u 
 
I N D N u m ber:  
1 3 1 3 5 3 
 
N C T N u m ber:   
N C T 0 2 8 0 7 5 9 7 
 
 
 
C O N FI D E N TI A L  
T he i nf or m ati o n c o nt ai ne d i n t his d oc u m e nt is re g ar de d as c o nfi de nti al a n d, e xce pt t o t he e xte nt necess ar y t o 
o bt ai n i nf or m e d c o nse nt, m a y n ot be discl ose d t o a n ot her p art y u nless l a w or re g ul ati o ns re q uire s uc h 
discl os ure.  Pers o ns t o w h o m t he i nf or m ati o n is discl ose d m ust be i nf or m e d t h at t he i nf or m ati o n is 
c o nfi de nti al a n d m a y n ot be f urt her discl ose d b y t he m  
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 of 5 4  
 
 
 
 
Si g n at ure P a ge 
 
 
T he si g nat ure bel o w c o nstit utes t he a p pr o val of t his pr ot oc ol a n d t he attac h me nts a n d pr o vi des t he 
necessar y ass ura n ces t hat t his st u d y will be c o n d ucte d acc or di n g t o all sti p ulati o ns of t he pr ot oc ol, 
i ncl u di n g all state me nts re gar di n g c o nfi de ntialit y, a n d acc or di n g t o l ocal le gal a n d re g ulat or y 
re q uire me nts a n d a p plica ble U. S. fe deral re g ulati o ns a n d I C H g ui deli nes. 
 
A m e n d m e nt/ Versi o n # _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
S T U - 2 0 2 2- 0 3 8 5 
E v al u ati o n of L S 3 0 1 U pt a ke i n T u m ors of P atie nts U n der g oi n g P arti al M astect o m y a n d 
Se nti nel L y m p h N o de Bi o ps y f or Bre ast C a ncer  
 
 
Pri nci p al I n vesti g at or ( PI) N a m e : _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
 
 
PI Si g n at ure: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
 
 
D ate: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 4 of 5 4  
 
 
S C H E M A  
 
 
 
L S 3 0 1 D ose Le vel  
L S 3 0 1 D ose  
Le vel 1 (starti n g d ose)  
0. 0 5 m g/ k g  
Le vel 2  
0. 0 7 5 m g/ k g  
Le vel 3  
0. 1 m g/ k g  
 
P hase 1 will be c o m plete after esta blis hi n g t he safet y a n d deter mi ni n g t he o pti mal i ma gi n g d ose 
of L S 3 0 1.  E nr oll me nt t o p hase 2 will o nl y be gi n after t he F D A h as re vi e w e d a n d a p pr o v e d t he 
p hase 1 data.
N e wl y dia g n ose d Sta ge I -II breast ca n cer patie nts u n der g oi n g breast-c o nser vi n g 
t hera p y a n d S L N  bi o ps y.  Ne gati ve n o dal b asi n o n cli nical e xa m. 
I njecti o n of L S 3 0 1 
( da y pri or t o s ur g er y if p ossi ble ( preferre d) or at l east 4 h o urs 
pri or t o s ur ger y o n d a y of s ur ger y) 
Patie nt m o nit ori n g f or 
o ne h o ur p ost-
i njecti o n 
3 0 -da y S O C la bs  
(f oll o w- u p) 
S ur ger y wit h Ca ncer Visi o n 
G o g gle S yste m  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 5 of 5 4  
 
 
T a ble of C o nte nts  
 
S C H E M A ....................................................................................................................................................... 4 
1. 0  
B A C K G R O U N D A N D R A TI O N A L E  .............................................................................................. 7 
1. 1  
S ur gi c al R e s e cti o n of Br e a st C a n c er a n d S e nti n el L y m p h N o d e Bi o p s y  .. 7 
1. 2  
I m a g e- G ui d e d N e ar-I nfr ar e d Fl u or e s c e n c e S y st e m s f or I ntr a o p er ati v e 
C a n c er I m a gi n g  ...................................................................................................................... 7 
1. 3  
W e ar a bl e C a n c er Vi si o n G o g gl e s f or S ur gi c al R e s e cti o n of C a n c er  ........... 8 
1. 4  
H u m a n Pil ot St u di e s U si n g W e ar a bl e C V G  ..................................................... 1 0 
1. 5  
NI R Fl u or e s c e nt M ol e c ul ar Pr o b e s f or t h e A c c ur at e I d e ntifi c ati o n of 
Di v er s e a n d H et er o g e n e o u s T u m or s u si n g NI R- FI G S I s L a c ki n g  ........................ 1 0 
1. 6  
NI R Fl u or e s c e nt M ol e c ul ar Pr o b e, L S 3 0 1, S el e cti v el y A c c u m ul at e s i n 
Br e a st C a n c er a n d Ot h er M ali g n a n ci e s  ....................................................................... 1 1 
1. 7  
M e c h a ni s m of L S 3 0 1 C a n c er- Bi n di n g A cti o n  ................................................. 1 1 
1. 8  
L S 3 0 1 Hi g hli g ht s T u m or s i n Di v er s e A ni m al M o d el s of C a n c er  ............... 1 3 
1. 9  
I N D- E n a bli n g Pr e cli ni c al St u di e s ........................................................................ 1 6 
1. 1 0  Cli ni c al St u d y Pl a n  .................................................................................................. 1 7 
1. 1 1  R ati o n al e  ..................................................................................................................... 1 8 
1. 1 2  P ote ntial  C o ntri b uti o n ............................................................................................... 1 8 
2. 0  
O B J E C TI V E S  ................................................................................................................................ 1 9  
2. 1  
Pri m ar y O bj e cti v e s  ................................................................................................... 1 9 
3. 0  
P A TI E N T S E L E C TI O N  ................................................................................................................ 1 9  
3. 1  
I n cl u si o n Crit eri a ...................................................................................................... 1 9 
3. 2  
E x cl u si o n Crit eri a  .................................................................................................... 1 9 
3. 3  
I n cl u si o n of W o m e n a n d Mi n oriti e s ................................................................... 1 9 
4. 0  
R E GI S T R A TI O N P R O C E D U R E S  ............................................................................................... 1 9  
4. 1  
C o nfir m ati o n of P ati e nt Eli gi bilit y  ....................................................................... 2 0 
4. 2  
P ati e nt R e gi str ati o n i n t h e Sit e m a n C a n c er C e nt er O n C or e D at a b a s e  
f or P h a s e 1 A & U T S o ut h w e st er n M e di c al C e nt er V el o s D at a b a s e f or P h a s e 
1 B / P h a s e 2  ............................................................................................................................ 2 0 
4. 3  
A s si g n m e nt of U P N  .................................................................................................. 2 0 
    4. 4     Baseli ne Data C ollecti o n  
5. 0  
S T U D Y P R O C E D U R E S ................................................................................................................ 2 0  
5. 1  
L S 3 0 1 Pr e p ar ati o n  ................................................................................................... 2 0 
5. 2  
Pr o c e d ur e s  .................................................................................................................. 2 1 
5. 3  
E v al u a bilit y  ................................................................................................................. 2 5 
5. 4  
S af et y E v al u ati o n  ..................................................................................................... 2 5 
5. 5  
Ri s k s a n d A d v er s e E v e nt M o nit ori n g  ................................................................. 2 7 
5. 6  
C oll e cti o n of S p e ci m e n s f or R e s e ar c h A n al y s e s ( P h a s e 2 O N L Y)  .............. 2 9 
5. 7  
D at a S u b mi s si o n S c h e d ul e  ................................................................................... 2 9 
5. 8  
W o m e n of C hil d b e ari n g P ot e nti al  ........................................................................ 3 0 
5. 9  
D ur ati o n of St u d y P arti ci p ati o n  .......................................................................... 3 0 
6. 0  
R E G U L A T O R Y A N D R E P O R TI N G R E Q UI R E M E N T S  ........................................................... 3 0  
6. 1  
S p o n s or-I n v e sti g at or R e p orti n g R e q uir e m e nt s – W a s hi n gt o n U ni v er sit y
  
3 1 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 6 of 5 4  
 
 
6. 2  
S p o n s or-I n v e sti g at or R e p orti n g R e q uir e m e nt s – U T S o ut h w e st er n 
M e di c al C e nt er  ...................................................................................................................... 3 3 
6. 3  
E x c e pti o n s t o E x p e dit e d R e p orti n g  .................................................................... 3 5 
7. 0  
D A T A A N D S A F E T Y M O NI T O RI N G  ......................................................................................... 3 5 
7. 1 P h a s e 1 A: W a s hi n g t o n U ni v e r si t y  ............................................................................................... 3 5  
7. 2  
P h a s e 1 B & P h a s e 2: U T  S o u t h w e s t e r n M e di c al C e n t e r ...................................................... 3 6  
8. 0  
S T A TI S TI C A L C O N SI D E R A TI O N S  ........................................................................................... 3 7 
8. 1  
St u d y D e si g n  .............................................................................................................. 3 7 
8. 2  
E n d p oi nt s  .................................................................................................................... 3 7 
8. 3  
D at a A n al y si s  ............................................................................................................. 3 8 
8. 4  
S a m pl e Si z e  ................................................................................................................ 3 9 
8. 5  
E arl y st o p pi n g r ul e  .................................................................................................. 4 0 
8. 6  
A n al y si s s et  ................................................................................................................ 4 0 
9. 0  
R E F E R E N C E S  .............................................................................................................................. 4 1  
1 0. 0  A P P E N DI X A:  D efi ni ti o n s f o r A d v e r s e E v e n t R e p o r ti n g  ................................................. 4 6 
1 1. 0  A P P E N DI X B:  R e p o r ti n g Ti m eli n e s  ....................................................................................... 4 8  
1 2. 0  A P P E N DI X C:  W a s hi n g t o n U ni v e r si t y U n a n ti ci p a t e d P r o bl e m R e p o r ti n g C o v e r 
S h e e t  5 2  
1 3. 0  A P P E N DI X D:  D S M C S e ri o u s A d v e r s e E v e n t R e p o r t F o r m  ............................................. 5 3  
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 7 of 5 4  
 
 
1. 0  
B A C K G R O U N D A N D R A TI O N A L E 
 
1. 1  
S ur gic al Resecti o n of B re ast C a ncer a n d Se nti nel L y m p h N o de Bi o ps y  
 
S ur gical res ecti o n is t he sta n dar d of care f or ma n y s oli d t u m ors s uc h as breast ca ncer, a n d 
se nti nel l y m p h n o de ( S L N) bi o ps y is use d f or ca ncer sta gi n g1. Bef ore breast s ur g er y, 
sta n dar d i ma gi n g met h o ds i ncl u di n g ma m m o gra p h y a n d ultras o u n d are use d t o i de ntif y 
t u m or l ocati o n. H o w e ver, t hese pre o perati ve i ma gi n g met h o ds are n ot a bl e t o pr o vi de real-
ti me i ma ge g ui da nce i n t he o perati n g r o o m t o e ns ure c o m plete re m o val of t u m ors or assess 
s ur gical mar gi ns. U nf ort u natel y, i nc o m plete t u m or re m o val i ncreases t he c ha nces of ca ncer 
rec urre nce b y  t w o -f ol d2 a n d  necessitates  re p eat  s ur ger y,  w hereas  i nacc urate  S L N  
i de ntificati o n ma y mis dia g n ose t he ca n cer sta ge. D es pite rece nt a d va n ces i n pre-o p erati ve 
i ma gi n g met h o ds, s ur ge o ns rel y o n vis ual i ns pecti o n, pal pati o n, a n d tactile e val uati o n t o 
disti n g uis h ca ncer fr o m u ni n v ol ve d tiss ue i ntra o perati vel y, lea di n g t o s u bjecti ve decisi o n-
ma ki n g a n d varia ble o ut c o mes. F or e x a m ple, u p t o 7 0 % 3- 5 of patie nts u n der g oi n g breast -
c o nser vi n g r ec o nstr u cti ve s ur ger y ( B CS) ha v e p ositi ve mar gi ns, de pe n di n g o n w here t h e 
s ur ger y wa s perf or me d, e x perie n ce of t he s ur ge o n, a n d t he t u m or c haract eristics6- 8.  F or 
t hese reas o ns, we see k t o de vel o p a n i ma ge-g ui de d met h o d f or differe ntiat i n g t u m ors fr o m 
s urr o u n di n g tiss ues a n d offer real-ti me s ur gi cal na vi gati o n 1 0 ,1 1. 
 
Si milarl y, assess me nt of t he S L N f or t he prese nce of ca ncer cells is sta n dar d of care f or 
sta gi n g  breast  ca ncer 1 ,1 2 ,1 3. I nacc urat e  S L N  re m o val  ofte n  re q uires  a d diti o nal  s ur gical  
i nter ve nti o ns1 4.  C o n ve nti o nall y,  patie nts  are  i njecte d  perit u m orall y  wit h  9 9 mTc  s ulf ur -
c oll oi d or a ne w C D 2 0 6 rece pt or -tar g ete d ra di o p h ar mace utical, 9 9 mTc Til ma n oce pt a n d a 
visi ble l y m p h otr o pic bl ue d ye. A ha n d h el d ga m ma -c o u nter is use d t o l o calize t he re gi o n of 
hi g hest ra di oacti vit y, a n d t he bl ue d y e ca n b e use d t o vis ualize t he S L N.  H o w e ver, 
ra di oacti ve S L N trac ki n g e x p oses p atie nts a n d h ealt h pr ofessi o nals t o i o nizi n g ra diati o n 
wit h o ut a direct S L N vis ualizati o n ca pa bilit y. Alt h o u g h bl ue d y es ca n b e vis ualize d wit h 
t he na ke d e ye, S L N tr ac ki n g b y t his met h o d is li mite d t o i ns pecti o n of o nl y s uperficial 
l y m p h  n o des  a n d  re q uir es  a  hi g h  d ose  of  t he  d ye,  w hic h  ma y  lea d  t o  har mf ul  si de 
effects 1 5 ,1 6. Wit h o ut k n o wi n g w hic h l y m p h n o des are p ositi ve, m ulti ple n o d es are re m o ve d 
ra n d o ml y f or s u bs e q ue nt hist ol o gic assess me nt. T h e a b o ve c halle n g es ha v e s p urre d i nterest 
i n de vel o pi n g met h o ds f or acc urate i ntra o perati v e detecti o n of t u m ors a n d S L Ns i n real-
ti me.  
 
1. 2  
I m a ge-G ui de d Ne ar -I nfr are d Fl u oresce nce S yste ms f or I ntr a o per ati ve 
C a ncer I m a gi n g  
 
T he pri mar y g oal of i ma ge g ui da nce i n t he o p erati n g r o o m is t o pr o vi de t h e s ur ge o n wit h 
acc ur ate, r eal-ti me i nf or mati o n a b o ut t he precise l ocati o n a n d b o u n dar y of t u m ors. T his 
will  als o  all o w  t he  s ur ge o n  t o  e x pl ore  alter nati ve  treat me nt  pla n ni n g  base d  o n  t he  
a g gressi ve ness of t he t u m or. A d va nce d i nstr u me nts t hat mi mic gl o bal p ositi o ni n g s yste ms 
ha ve als o bee n de vel o p e d, w here a pre o p erati ve C T or M R i ma ge ca n be pr ojecte d o nt o 
t he a p pr o priate a nat o mical str uct ures. T hese s yste ms s uffer fr o m t he li mitati o ns of t he pre -
o perati ve  i ma gi n g  met h o d,  u nsatisfact or y  re gistrati o n  d ue  t o  tiss ue  def or mati o n  a n d  
m oti o n, a n d lac k of t he a bilit y t o i ntra o perati vel y i nterr o gate s ur gical mar gi ns f or t he 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 8 of 5 4  
 
 
prese nce of t u m ors.  
 
R ece nt a d v a nces i n bi o p h ot o nics a n d se mic o n d u ct or tec h n ol o gies h a ve accelerate d t h e 
e mer ge n ce of o ptical i ma gi n g as a para di g m -s hifti n g met h o d f or real-ti me ca ncer detecti o n. 
O ptical i ma gi n g ca n det ect bi ol o gical e ve nts ra n gi n g fr o m m olec ular a n d s u b -cell ular 
le vels t o t he le vel of e ntire or ga n s yst e m s. I n t he near i nfrare d ( NI R ) re gi o n bet wee n 7 0 0 
a n d 9 0 0 n m, a bs or pti o n b y i ntri nsic p h ot oacti ve bi o m olec ules is l o w, w hic h mi ni mizes 
tiss ue a ut ofl u oresce n ce a n d facilitates t hic k tiss ue assess me nt2 1. U nli ke n uclear met h o ds, 
o ptical i ma gi n g utilizes n o ni o nizi n g ra diati o n. T his e x plai ns t he rece nt s ur g e i n i nterest i n 
a p pl yi n g t he met h o d i n i ntra o perati ve pr o ce d ures, w here t u m or b o u n daries, l y m p h n o de 
assess me nt, treat me nt res p o nse, a n d i m p orta nt p h ysi ol o gical para meters ca n be m o nit ore d 
i n real-ti me. T o date, vari o us o ptical tec h n ol o gies s uc h as fl u oresce nce i ma gi n g ha ve bee n 
a p plie d t o assess t u m or m ar gi ns 2 2 -3 1. Nearl y all t he s yste ms utilize t he NI R li g ht, w hic h ca n 
detect fl u or esce nt o bjects u p t o 1 c m d ee p i n s oli d tiss ue a n d 5 c m i n l u n g tiss ue. T hese 
o ptical i ma gi n g de vices offer hi g h -res ol uti o n (t y picall y 1 2 5 – 6 2 5 µ m) a n d lar ge fiel d s-of -
vie w ( 7 - 1 5 c m).  
 
T o da te, s evera l NIR-f l u oresce nce i ma ge - g uide d s u r ger y ( NI R- FI G S) s yste ms ha ve be en 
d e vel o pe d a n d s uccessf ully use d f or  i ntra o perative t u m or ima gi n g a n d S L N   ma p pi n g 
( S L N M), incl u di n g F L A R E 3 2, Fl uo bea m 3 3, S P Y 3 4, Pal o mar, a n d H a ma mat s u P D E3 5. T he 
S p y ™ s yste m fr o m N o va da q uses a n NI R laser t o e xcite t he s ur gical fiel d. T he S p y ™ 
s yste m has alrea d y b ee n gra nte d t hree se parate 5 1 0( k) a p pr o v als.  
 
1. 3  
We ar a ble C a ncer Visi o n G o g gles f or S ur gic al R esecti o n of C a ncer  
 
M ost  of  t he  c urre nt  o ptical  i ma gi n g  s yste ms  use d  i n  t he  NI R-FI G S  ha ve  m aj or  
s h ortc o mi n gs t hat i m pe de t he ra pi d a d o pti o n a n d sea mless i nte grati o n i nt o t he w or kfl o w 
of o nc ol o gic s ur ger y. T hese li mitati o ns i ncl u de t he use   of b ul k y har d w are, p ote ntiall y 
disr u pti ve i nf or mati o n dis pl a y o n a re mote m o nit or, m ismat c h betwee n t he  s ystem ’s a n d 
s ur ge o n’s fiel d of view ( F O V ), a d diti o nal eff ort fr o m a n ot her s ur gical staff t o c o ntr ol t he 
de v ice,  hi g h c ost , a n d c o m ple x i nstr u me ntati o n. T o o verc o me t hese li mitati o ns, our tea m 
rece ntl y  de vel o pe d a n d teste d  i n ca ncer patie nts  a ne w hea d -m o u nte d i ma gi n g s yste m, 
Ca ncer Visi o n G o g gles  ( C V G), f or use i n t he o perati n g r o o m3 6 -4 3. T he sc he matics a n d 
pr ot ot y pes of t he C V G s yste m are s h o w n i n Fi g ure s 1 a n d 2.    
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 9 of 5 4  
 
 
 
Fi g ure 1: Sc he matics of t he Ca ncer Visi o n G o g gles s yste m. F P G A - fiel d- pr o gra m ma ble 
gate arr a y; H D MI - hi g h-defi niti o n m ulti me dia i nterface; NI R – near -i nfrare d li g ht; R G B 
– re d, bl ue, gr ee n c ol or s pace; P C – pr ocessi n g c o m p uter u nit 4 3.   
 
 
Fi g ure 2: Pr ot ot y pes of t he Ca ncer Visi o n G o g gles s yste ms. (a) Vi de o -s ee-t hr o u g h versi o n 
w or n b y Dr. J ulie Mar g e nt haler i n t he o perati n g r o o m. ( B) O ptical -see-t hr o u g h versi o n 
w or n b y a r esearc her. B ot h s yste ms ha ve b ee n use d i n t he o p erati n g r o o m f or breast -
c o nser vi n g s ur ger y b y Dr. Mar ge nt haler 4 3. 
 
T he C V G h ol ds great pr o mise t o o verc o m e t he c hallen ges of t he  c urre nt NI R -FI G S  s yste ms 
a n d  e x pa n d  fl u ores ce nce -g ui de d  s ur ger y  i nt o  a d va nce d  cli nical  ce nters  as  well  as  
u n derser v e d areas, es peciall y t h ose i n l o w -res o urce setti n gs. T h is si m ple g o g gle-base d 
de vice is aff or da bl e, c o m pact, weara ble, a n d all o ws ha n ds -free s ur gi cal o perati o n. Wit h 
t his de vice, f u n cti o nal i nf or mati o n pr o vi de d b y t he NI R fl u oresce n ce of c o ntrast a ge nts i n 
ca ncer  cells  or  t he t u m or  e n vir o n me nt  ca n  be  directl y  dis pla ye d  wit h  a dj usta ble  
a m plificati o n o n t he g o g gles’ e ye piece. T h e C V G s yste m pr ojects b ot h NI R  fl u oresce n ce 
fr o m t u m ors a n d t he nat ural c ol or i ma ges of tiss ue o nt o a hea d m o u nte d dis pla y wit h o ut 
late nc y4 3.  
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 0  of 5 4  
 
 
1. 4  
H u m a n Pil ot St u dies U si n g We ar a ble C V G  
 
Cli nical feasi bilit y was d e m o nstrate d i n 1 5 breast ca ncer a n d 5 m ela n o ma patie nts d uri n g 
S L N   bi o ps y after l u m pect o m y  or mastect o m y f or breast or wi de e xcisi o n s ur geries  f or 
mela n o ma .  I n d oc ya ni n e  gree n  (I C G),  a n  F D A-a p pr o ve d  NI R  fl u oresce nt  d ye,  was  
e m pl o ye d as t he i ma gi n g a ge nt. S ur ge o ns use d t he i ma gi n g s yste m c o mf orta bl y wit h 
mi ni mal disr u pti o n t o t he s ur gical w or kfl o w. T he C V G all o we d clear vis ualizati o n of 3 0 
S L Ns fr o m 1 0 breast ca n cer a n d 5 mel a n o ma patie nts. Usi n g hist ol o gic a nal ysis as t he g ol d 
sta n dar d, t he C V G ha d a detecti o n se nsiti vit y of 1 0 0 % i n c o m paris o n t o 9 2. 8 6 % ± 1 7. 5 % 
f or t he bl ue d ye a n d 9 6. 4 3 % ± 1 2. 9 % f or ra di oacti ve trac ki n g. I n o ne mela n o ma patie nt 
( Fi g. 6), bl u e d ye di d n ot i de ntif y t w o dee p-seate d S L Ns. Si milarl y, i n o ne breast ca n cer 
patie nt, i nitial vis ual i ns pecti o n di d n ot re veal t he S L N a n d i n a n ot her patie nt, ra di oacti ve 
trac ki n g was u na bl e t o i de ntif y t w o S L Ns. I n t hese cases, t he S L Ns were clearl y i de ntifie d 
b y C V G. Alt h o u g h t he i ma gi n g de pt h wit h reas o na ble res ol uti o n is a b o ut 5 m m, hi g h 
fl u oresce n ce si g nal fr o m dee p-seat e d S L Ns is rea dil y pr ojecte d t o t he s urface, all o wi n g 
vis ualizati o n of S L Ns at > 5 m m dee p after defl ecti o n of t he o verl yi n g tiss ue la yer. T his 
pil ot st u d y de m o nstrates t he p ote ntial cli nical utilit y of t he s yste m f or ra pi d i de ntificati o n 
of S L Ns d uri n g s ur ger y. 
 
B y F D A g ui deli nes, t he C V G de vice is c o nsi dere d a “ n o n-si g nifica nt ris k” ( N S R) de vice 
a n d d oes n ot c o me i nt o direct c o ntact wit h patie nt tiss ue d uri n g s ur ger y. We will use t hese 
s yste ms t o assess t he NI R fl u oresce nce of t u m or-tar geti n g o ptical c o ntrast a ge nts i n t he 
pla n ne d st u d y. 
 
1. 5  
NI R Fl u oresce nt M olec ul ar Pr o bes f or t he A cc ur ate I de ntific ati o n of 
Di verse a n d Heter o ge n eo us T u m ors usi n g NI R -FI G S Is L ac ki n g 
 
M ost NI R - FI G S s yste ms are desi g ne d t o o perate i n t he NI R wa v ele n gt hs. S uccessf ul 
i m ple me ntati o n of real-ti me NI R -FI G S  f or m ore acc urate t u m or resecti o n re q uires t he 
de vel o p me nt of NI R fl u oresce nt i ma gi n g a ge nts t h at ca n s electi vel y bi n d or acc u m ulate i n 
heter o ge n e o us ca ncer  cells ,  i n de pe n de nt  of  t u m or  t y pe. Clearl y,  t he  de vel o p me nt  of  
acc ur ate a n d br oa d ca n cer -s pecifi c i ma gi n g a ge nts will accelerate cli nical tra nslati o n a n d 
use of o ptical i ma gi n g m et h o ds i n cli nics . O ptical m olec ular  i ma gi n g has e v ol ve d fr o m a 
pre d o mi na ntl y precli nical i ma gi n g tec h n ol o g y t o i ncreasi n g a p plicati o n s i n cli nics. T he 
pr o mise of hi g h detecti o n se nsiti vit y a n d real-ti me i ma gi n g fee d b ac k has res ulte d i n t he 
de vel o p me nt of a m yria d of o ptical m olec ular i ma gi n g pr o bes f or ca ncer a p plicati o n, 
ra n gi n g  fr o m  r ece pt or-me diate d  t o  e nz y m e  acti vata ble  m olec ular  pr o b es. 4 4 ,4 5 N ota ble 
e xa m ples i ncl u de t he use of fl u oresce nt i ma gi n g a ge nts i n l y m p h n o de ma p pi n g, i ma ge -
g ui de d  s ur ger y,  o p ht hal m ol o gic  a p plicati o ns,  a n d  e val uati o n of  l y m p h atic  fl o w. 
Researc h ers are acti vel y p urs ui n g t w o maj or re p orti n g strate gies i n de vel o pi n g m olec ular 
i ma gi n g pr o bes f or h u m a n use. First, a ffi nit y-bas e d  m olec ular pr o b es are c o ntrast a g e nts 
t hat selecti vel y acc u m ul ate i n t u m or tiss ue relati ve t o t he s urr o u n di n g n or mal tiss ue b y 
bi n di n g t o o ver -e x press e d pr otei ns or t hr o u g h ot her u pta k e mec ha nis ms. Affi nit y pr o bes 
t y picall y i n v ol ve t he c o nj u gati o n of a fl u oresce nt d ye t o t u m or-tar geti n g bi o m olec ules, 
s uc h as m o n ocl o nal a nti b o dies or hi g h affi nit y p e pti de li ga n ds. H o we ver, t his met h o d is 
c o m plicate d  b y  i ntri nsic  e x pressi o n  of  tar gete d  pr otei ns  i n  healt h y  tiss ues,  t u m or  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 1  of 5 4  
 
 
heter o ge n eit y,  a n d  patie nt -de pe n d e nt  varia bilit y. 4 6 -5 3  A  sec o n d a p pr oac h  is  t o  use  
acti vata ble fl u or esce nt m olec ular pr o bes.  T hes e m olec ular pr o bes are d esi g ne d t o ha ve 
l o w fl u oresce nce yiel d u ntil t he y e nc o u nter a m olec ular tar get (e. g. e n z y me acti vata ble 
pr o bes) 5 4 or l ocalize i n f a v ora ble p h ysi ol o gical m e di u m (e. g. p H acti vata ble pr o bes). 5 5 ,5 6 
Alt h o u g h t hese pr o bes h a ve l o w b ac k gr o u n d fl u oresce n ce, t h e p ol y meric materials use d 
f or t heir de v el o p me nt res ults i n ver y sl o w fl u oresce nce e n ha n ce me nt, re q uiri n g se veral 
h o urs f or o pti mal si g nal e n ha nce me nt i n t u m ors, a n d ma y be c o m plicate d b y n o n -s pecifi c 
acti vati o n. F urt her, s yste mic a d mi nistrati o n of s u c h macr o m olec ules ca n elicit reacti o ns i n 
patie nts, a ris k t hat ma y li mit t heir use.  
 
W hat is nee d e d t o har n ess t he f ull p ote ntial of o ptical i ma gi n g (si m plicit y, u n parallele d 
detecti o n se nsiti vit y, a n d real-ti me fee d bac k) is t he de vel o p me nt of a NI R m olec ular pr o b e 
t hat ca n selecti vel y acc u m ulate i n ca ncer cells wit h hi g h s pecificit y, a n d a m olec ule wit h 
wi de a p plicati o ns si milar t o 2- de o x y- 2- [fl u ori n e-1 8]fl u or o - D-gl uc ose ( 1 8F- F D G), w hic h 
re v ol uti o nize d ca n cer dia g n osis t hr o u g h t he use of P E T.  
 
1. 6  
NI R Fl u oresce nt M olec ul ar Pr o be, L S 3 0 1, Selecti vel y Acc u m ul ates i n  Bre ast 
C a ncer a n d Ot her M ali g n a ncies  
 
We ha ve c h a m pi o ne d t he de vel o p me nt of di vers e NI R fl u oresce nt d ye -la bele d pe pti des 
(affi nit y pr o b es) f or use i n t he m olec ular o ptical i ma gi n g of t u m ors, as e vi de nce d b y t h e 
first de m o nstrati o n of t his a p pr oac h i n r o de nts5 7 ,5 8. As n ote d a b o ve, ca n cer heter o ge n eit y, 
e ve n wit hi n t he sa m e ca ncer p atie nt, c o nfi nes t he use of t hese i ma gi n g a ge nts t o o nl y 
s pecific ca n cer s u b-t y p es t hat o vere x press t he tar g et bi ol o gical bi o mar kers. We ha ve n o w 
disc o vere d a n e w m olec ular pr o be, L S 3 0 1, w hic h selecti vel y acc u m ulates i n pri mar y breast 
a n d ot her ca ncers (es p eci all y at t he t u m or peri p her y), s yste mic metastases a n d metastatic 
l y m p h n o des.  
 
1. 7  
Mec h a nis m of L S 3 0 1 C a ncer- Bi n di n g Acti o n  
 
L S 3 0 1 ( Fi g ure 3 ) is a s mall m olec ule ( < 1. 6 k D a) c o nsisti n g of a NI R fl u oresce nt d ye 
(c y pate)5 9 ,6 0 a n d a n octa pe pti de t hat is c y clize d t hr o u g h a dis ulfi de b o n d. T he NI R d ye, 
i n d oc ya ni ne gr ee n (I C G), is a n F D A-a p pr o ve d d y e f or use i n h u m a n s u bjects. T he s pectral 
pr o perties of I C G are s uita ble f or NI R -FI G S (e x citati o n/e missi o n 7 8 5/ 8 1 0 n m i n ser u m 
a n d a s ol uti o n of 2 0 % D M S O i n a n a q u e o us me di u m; m olar a bs or pti vit y, ε, 2 4 0, 0 0 0 M -
1.c m- 1;  a n d  fl u oresce nce  q ua nt u m  yiel d,  ψ,  1 0 %).  T his  d ye  has  bee n  e m pl o ye d  t o 
de m o nstrate  t he  feasi bilit y  of  usi n g  o ptical  i ma gi n g  met h o ds  t o  st u d y  h u ma n  
pat h o p h ysi ol o gical c o n diti o ns. U nf ort u natel y, I C G is n ot t u m or -selecti ve a n d its i na bilit y 
t o react wit h bi o m olec ul es has pre v e nte d its use f or la beli n g t u m or-tar g eti n g bi ol o gical 
carri ers 6 1 ,6 2. T o a d dress t his nee d, w e d e vel o pe d a n I C G deri vati ve, c y p ate, w hic h has 
si milar bi ol o gical cleara n ce pr ofile a n d s pectral pr o perties (a bs or pti o n, e missi o n, q ua nt u m 
yiel d) t o I C G. C y pate bi n ds re versi bl y t o t he h y dr o p h o bic p oc ket of al b u mi n, 6 3 ,6 4 a s o urce 
of nitr o ge n a n d e n er g y f or t u m ors. 6 5 -6 7 U nli ke c o vale ntl y d ye -la bel e d al b u mi n m olec ular 
pr o bes, c y pate is r elease d i n t u m ors u n der mil d aci dic c o n diti o ns as it traffics t hr o u g h t he 
e n d os o mal pat h wa y. T h e releas e d c y pate is o nl y tra nsie ntl y retai ne d i n t u m ors bef ore 
effl u x, li miti n g t he t u m or -t o- bac k gr o u n d c o ntrast.  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 2  of 5 4  
 
 
 
 
Fi g ure 3: Str uct ure ( Left) a n d s pectral pr o perties of L S 3 0 1 ( Ri g ht). 
 
T o i m pr o ve t u m or rete nti o n, we e x pl ore d usi n g p e pti des t o tra p t he i nter n alize d c y pat e. 
T he hi g hl y re d uci n g e n vir o n me nt of t u m or cells is k n o w n t o re d uce dis ulfi de b o n ds t o 
t hi ols, w hic h ca n tra ns-t hi olate wit h c ystei ne-c o nt ai ni n g i ntracell ular pr otei ns a n d tra p t he 
m olec ular pr o be  i n ca ncer cells . T heref ore, we c o nj u gate d a variet y of dis ulfi de-c o ntai ni n g 
pe pti des t o c y pate a n d s cree n e d t he c o m p o u n ds i n i m m ortalize d h u ma n ca ncer cell li nes 
a n d mice wit h di vers e ca ncer xe n o grafts . O ur res ults s h o we d t hat L S 3 0 1 was selecti vel y 
retai ne d  i n  t u m or tiss ues c o m pare d  t o  t he  ot her  pe pti de  c o nj u gates.  Str uct ure-t u m or 
rete nti o n a nal ysis re v eal e d t hat t he u n nat ural D -cystei ne li n ke d t o c y pat e c o nfers hi g h 
bi ol o gical sta bilit y o n t he m olec ular pr o be b eca us e of its resista nce t o ra pi d de gra dati o n b y 
p r oteases. 
 
NI R  fl u ores ce nce  micr osc o p y  of  L S 3 0 1  i n  di verse  t u m or  cells  s h o we d  p u nct ate  
i ntracell ular fl u ores ce nce t y pical of rece pt or-me diate d e n d oc yt osis ( Fi g ure 4 ). Wester n 
bl ot, pr ote o mic a nal ysis, a n d bl oc ki n g st u dies i n cells wit h di verse i n hi bit ors of al b u mi n 
e n d oc yt osis p oi nt t o a n i nitial al b u mi n -me diate d e n d oc yt osis. T his mec ha nis m of u pta ke 
was f urt her s u p p orte d i n vitr o a n d i n viv o b y c o -i nc u bati o n of Ale xa 6 8 0 l a bele d b o vi n e 
ser u m al b u mi n ( B S A) wit h L S 3 0 1, w hic h s h o we d c o-l ocalizati o n of t he t w o fl u or o p h ores 
i n ca ncer cells at earl y ti me p oi nts. Di ver ge nce of t he d ye si g nals was o bser ve d at later 
ti me-p oi nts, i n dicati n g t he effl u x of t he Ale xa 6 8 0 fr o m t he ca ncer cells, w hile L S 3 0 1 
fl u oresce n ce w as retai ne d. Calci u m als o accelerates i nter nalizati o n of L S 3 0 1 i n ca ncer 
cells a n d a d diti o nal cell st u dies i n dicate t h at o ne of t he i ntracell ular pr otei ns t hat bi n d t his 
m olec ular pr o b e is p h os p h or ylate d  A n n e xi n A 2  ( p A n x A 2), a n i n dicat or of t he c hr o nic 
i nfla m mat or y e n vir o n me nt of t u m ors. Base d o n t he o n g oi n g st u dies, part of t he mec ha nis m 
of L S 3 0 1 u pta ke a n d r ete nti o n i n ca ncer cells i n v ol ves al b u mi n -facilitate d tra ns p ort t o 
t u m ors, f oll o we d b y bi n di n g t o p A n x A 2 o n t u m or cells a n d t u m or-ass ociat e d str o mal cells. 
T he  c o m ple x  traffics  i nt o  t he  cells  w here  L S 3 0 1  ca n  be  f urt her  sta bilize d  via  tra ns-
t hi olati o n  of  t he  D-c ystei ne wit h  i ntracell ular  pr otei ns  u n der  t he  hi g hl y  re d uci n g  
e n vir o n me nt of ca ncer cells. T he pe pti de is e ve nt uall y de gra de d a n d cleare d fr o m t he b o d y 
a n d t he d ye is cleare d vi a he pat o biliar y e xcreti o n pat h wa y si milar t o I C G.  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 3  of 5 4  
 
 
 
Fi g ure 4: I nter nalizati o n of L S 3 0 1 i n breast ca n cer ( 4 T 1cells. Re d: L S 3 0; Bl ue: n uclear 
stai n ( D A PI). 
 
1. 8  
L S 3 0 1 Hi g hli g hts T u m ors i n Di verse A ni m al M o dels of C a ncer  
 
L S 3 0 1 is pr o d uce d at t he O ptical Ra di ol o g y L a b orat or y  at Was hi n gt o n U ni versit y Sc h o ol 
of Me dici ne i n c o nf or ma nce wit h G o o d L a b orat or y Practices as descri be d i n C F R 2 1 Parts 
2 1 0 a n d 2 1 1.  We ha v e e v al uate d L S 3 0 1 i n di verse t u m or m o dels i n mice, i ncl u di n g breast, 
l u n g,  c ol o n,  li ver,  pr ostate,  pa ncreatic,  a n d  brai n  t u m ors.  I n  all  cas es,  L S 3 0 1  was  
selecti vel y retai n e d i n o nl y t he t u m or tiss ue f or all differe nt t u m or t y pes ( Fi g ure 5 ).  
 
 
Fi g ure 5: Re pr ese ntati v e pla nar i ma ges of t u m or m o dels t hat selecti vel y retai n L S 3 0 1. 
Ort h ot o pic 4 T 1l uc m uri ne ( Left), D B T brai n ( Mi d dle), a n d H T 2 9 c ol o n ( Ri g ht) ca ncer 
m o dels. False c ol or f or L S 3 0 1 fl u oresce nce is re d  i n t he first t w o pa n els a n d rai n b o w i n 
t he last pa nel wit h bl ue as l o west i nte nsit y a n d re d as hi g hest i nte nsit y o btai ne d 2 4 h o urs 
after i njecti o n of L S 3 0 1.    
 
We  ha v e  als o  de m o nstrate d  t hat  L S 3 0 1  ca n  disti n g uis h  i nfla m mati o n  fr o m  c ol o n  
a de n ocarci n o ma i n c olitis m o del of c ol o n ca ncer , as w ell as p a ncreatitis fr o m pa ncreatic 
ca ncer.  T he  i ma gi n g  a g e nt  als o  i de ntifies  s p o nta ne o us  micr osc o pic  breast  t u m ors  or  
disse mi nate d cer v ical t u m ors wit h hi g h acc urac y, as well as p ositi ve l y m p h n o des (Fi g ure 
6 ).  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 4  of 5 4  
 
 
 
Fi g ure 6:  L S 3 0 1 i de ntifies  l y m p h n o des i nfiltrate d b y met astatic breast  ca ncer cells. 
Pre o perati ve ( Left) a n d after re m o vi n g t he s ki n. Gree n: L S 3 0 1 fl u ores ce nce i n p ositi ve 
l y m p h n o des. 
 
T o si m ulate h u ma n breast ca ncer a p plicati o n, we teste d L S 3 0 1 i n tri ple -ne gati ve breast 
ca ncer usi n g t he cli nicall y-rele v a nt patie nt- deri v e d xe n o graft ( P D X) m o dels (Fi g ure 7 ).   
 
 
Fi g ure 7:  L S 3 0 1 u pta ke i n P D X breast ca n cer m o del. Re d: L S 3 0 1 fl u oresce nce. 
 
1. 8. 1  Preli mi n ar y ( N o n -G L P) A ni m al T o xicit y Testi n g  i n Mice  
 
P ote ntial t o xicit y of L S 3 0 1 has bee n test e d i n 6 0 y o u n g a d ult male a n d fe male C D -
1 I G S mice  wit h a n a v era ge b o d y wei g ht of 2 0 g .  T w e nt y males a n d 2 0 fe males 
were eac h a d mi nistere d a si n gle d ose of 7. 5 m g/ k g L S 3 0 1 ( 1 5 0 µ g i n 1 0 0 µ L sali ne 
c o ntai ni n g 1 % m uri ne ser u m al b u mi n ) i ntra ve n o usl y via t he lateral tail vei n. Te n 
c o ntr ols of eac h ge n der w ere a d mi nistere d a si n gle d ose of 1 0 0 µ L sali ne. Te n male 
a n d 1 0 fe m ale dr u g -treat e d a ni mals a n d 5 male a n d 5 fe male c o ntr ol a ni mals were 
e ut ha nize d  2  da ys  p ost -L S 3 0 1  a d mi nistrati o n .   T he  re mai ni n g  a ni mals  were  
e ut ha nize d  1 4 d a ys p ost- L S 3 0 1 a d mi nistrati o n. 
 
T here was n o i nci de nce of u ne x pecte d deat h. All a ni mals s ur vi ve d a n d re mai ne d 
healt h y u ntil t he pla n ne d e n d p oi nts. N o cli nical a b n or malities were n ot e d i n a n y of 
t he a ni mals. T her e w ere n o differe nces bet wee n t he treat e d a n d c o ntr ol gr o u ps f or 
all meas ure d he m at ol o g y para meters f or b ot h t he D a y 2 a n d Da y 1 4 ti me p oi nts.  
T he s o di u m a n d c hl ori d e le vels of treat e d male a ni mals e val uate d o n Da y 2 were 
sli g htl y l o wer t ha n t h ose of t he c o m para ble c o ntr ol a ni mals, b ut were n or mal b y 
Da y 1 4. T her e were n o a b n or mal gr oss necr o ps y fi n di n gs attri b uta ble t o L S 3 0 1. 
 
We f o u n d t hat  micr o gra n ul o mata  f or me d ar o u n d g ol de n-br o w n f oci of pi g me nt i n 
t he l u n gs.  T his fi n di n g was o bser ve d i n 1 6 of t h e 2 0 treate d a ni mals e val uate d o n 
Da y 2, wit h b ot h g e n ders si milarl y affecte d, a n d was n ot f o u n d i n a n y c o ntr ol 
a ni mals.  S o me of t hese f oci were f o u n d  wit hi n vessel l u me ns, b ut n o f urt her t ha n 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 5  of 5 4  
 
 
t he al ve olar wall.  T hese lesi o ns were a bse nt i n t he Da y 1 4 a ni mals, s u g gesti n g 
res ol uti o n of t his lesi o ns wit hi n t he 2 -wee k ti me fra me.  
 
1. 8. 2  Preli mi n ar y ( N o n -G L P) T o xicit y Testi n g i n C a ni nes 
 
T hree 4 - m o nt h-ol d fe mal e d o gs were treate d wit h 0. 9 m g/ k g L S 3 0 1 i ntra ve n o usl y 
o ver 5 mi n utes.  A ni mals were m o nit ore d cl osel y f or t he first h o ur f or c h a n ges i n 
be ha vi or, r es pirati o n, heart rate, a n d bl o o d press ure, t he n a gai n at 4, 2 4 a n d 4 8 h o urs 
p ost i njecti o n (see attac he d t o xicit y re p ort). Bl o o d was dra w n f or he mat ol o g y a n d 
cli nical c he mistries bef ore ( baseli ne) a n d at 1, 4, 2 4, 4 8 a n d 7 da ys after i njecti o n. 
O ne of t he d o gs v o mite d s o o n after i nf usi o n wit h o ut a n y ot her a d vers e effects. All 
a ni mals  be ha v e d  n or m all y  w he n  release d  a n d  c o nti n ue d  n or m al  b e ha vi or  
t hr o u g h o ut t he d urati o n of t he trial.  
 
T he w h ole bl o o d was ce ntrif u ge d at 3 0 0 0  x g  f or 1 0 mi nutes a n d  t he plas ma was 
se parat e d fr o m re d bl o o d cells wit h pi pettes. T he plas ma sa m ples at differe nt ti me 
p oi nt were meas ure d o n Bec k ma n C o ulter D U 6 4 0 s pectr o p h ot o meter directl y 
usi n g u n -treate d plas ma sa m ples as baseli nes  (Fi g ure 8 ). T he c o nce ntrati o ns of t he 
pr o be wer e cal c ulate d usi n g t he  Beer – La m bert la w a n d t he e xti ncti o n c oefficie nt 
of t he pr o be L S 3 0 1. O ne -p hase e x p o n e ntial fitti n g was perf or me d t o det er mi ne t he 
half -life. 
 
Fi g ure 8:  L S 3 0 1 c o n ce ntrati o ns i n serial plas ma sa m ples deter mi ne d b y a bs or pti o n 
s pectr osc o p y wit h pre-i nj ecti o n plas ma val ues as baseli ne. O ne-p h ase e x p o ne ntial 
least-s q uar es fitti n g deter mi ne d a n a vera g e eli mi nati o n half-life of 4. 8 +/- 0. 5 h o ur 
(T a ble 1 ). 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 6  of 5 4  
 
 
T a ble 1:  Calc ulate d bl o o d cleara nce para meters f or L S 3 0 1 i n d o gs. 
P ar a m eter  
D o g 5 2 0 8  
D o g 5 2 1 4  
D o g 5 2 1 5  
K  
0. 0 0 2 1 9 0  
0. 0 0 2 7 4 5  
0. 0 0 2 3 7 8  
Half Life  
3 1 6. 5  
2 5 2. 5  
2 9 1. 5  
R²  
0. 9 7 3 5  
0. 9 7 5 4  
0. 9 7 9 6  
 
O ne of t he pri mar y fi n di n gs was n ote d i n t he p eri p heral bl o o d of d o g 5 2 1 4 i n t he 
i nitial  p ost-treat me nt  peri o d.   T his  a ni mal  s h o we d  a  dr o p  i n  ne utr o p hils  fr o m  
baseli ne le vels o v er t he c o urse of 5 -6 0 mi n utes p ost -dr u g a d mi nistrati o n, wit h 
rec o ver y of t hese ne utr o p hil n u m bers 4 h o urs p ost-treat me nt.  T his c ha n ge was 
n ota ble, a n d ma y ha v e s u g geste d eit her mar gi nati o n of ne utr o p hils, or a s hift of 
ne utr o p hils t o t he or ga ns.  Base d o n o ur c o ns ultati o n wit h a veteri n ar y cli nical 
pat h ol o gist, t hese fi n di n gs ma y res ult fr o m eit her e n d ot o xe mia or i di os y ncratic 
dr u g reacti o n; platelets ma y f oll o w a s a me patter n as t he n e utr o p hils.  T he ot her 
a ni mals di d n ot s h o w a si milar patter n i n t hese n u m bers.  Gi ve n e n d ot o xe mia w o ul d 
be e x p ecte d t o ha ve affecte d all a ni mals, t his  fi n di n g was pres u me d t o reflect a n 
i di os y ncratic dr u g r eacti o n i n d o g 5 2 1 4.   
 
A n ot her fi n di n g i n t he p eri p heral bl o o d t hat was n ote d i n all t hree a ni mals was a 
sli g ht ele vati o n i n cr eati ni ne le vels at t he 4 h o ur a n d 2 4 h o ur ti me p oi nts (t his 
a nal yte w as n ot meas ure d at earlier ti me p oi nts).  F oll o wi n g f urt her i n v esti gati o n, 
it was deter mi ne d t h at t his li kel y reflecte d a n i nteracti o n of t he test c o m p o u n d wit h 
picric aci d use d as p art of t he test met h o d ol o g y  f or meas uri n g creati ni ne, f or mi n g 
a c hr o m o ge n t hat mi mic ke d t he a bs or b a nce e x pecte d f or creati ni ne. 
 
Asi de fr o m t he p ul m o n ar y c o n gesti o n a n d e d e ma alrea d y dis c usse d a b o ve, t he 
pri mar y fi n di n gs i n t he l u n gs i ncl u de d mi ni mal t o mil d s mall f oci of p ne u m o nitis.  
T hese wer e of li mite d distri b uti o n.  T he ca use a n d si g nifica nce of t hese, al o n g wit h 
t he s mall f oci of i nfla m mati o n n ote d i n t he li vers of all d o gs is n ot k n o w n; h o we ver, 
t hese are c o nsi dere d c o m m o n fi n di n gs i n y o u n g d o gs, a n d are t h o u g ht t o b e rel ate d 
t o pri or parasitic i nfecti o n.  I n partic ular, i nfecti o n wit h Ascari ds earl y i n life ca n 
pr o d uce s uc h lesi o ns.  C o m paris o n wit h c o ntr ol a ni mals o n a f ut ure st u d y ma y hel p 
clarif y  t his   o bser vati o n.   Als o  n ote d  i n  all  t hree  d o gs  were  s mall  f oci  of  
mi neralizati o n i n t he re n al pa pillae.  T h ese are ver y c o m m o n fi n di n gs i n d o gs a n d 
were n ot li kel y a tr eat me nt effect.  
 
1. 9  
I N D-E n a bli n g Precli nic al St u dies  
 
L S 3 0 1 was ma n ufact ur e d as re q uire d b y t he C urre nt G o o d Ma n ufact uri n g Practice (c G M P) 
Re g ulati o ns a n d use d i n safet y st u dies rec o m me n de d b y t he A ge nc y t o s u p p ort cli nical 
i n vesti gati o n of L S 3 0 1. A n a de q uatel y-d esi g ne d a n d c o ntr olle d safet y p har mac ol o g y st u d y 
i n  d o gs,  a n  a de q uatel y  desi g ne d  si n gle-d ose  t o xicit y  st u d y  i n  rats,  a n d  a  bacterial  
m uta ge nicit y te st ( A mes assa y) w ere c o n d ucte d i n c o m plia nce wit h G L P re g ulati o ns, a n d 
t he st u d y re p orts c o ntai n all re q uire d i nf or mati o n f or G L P c o m plia nce. T he test s yste ms 
use d i n t he st u dies were a p pr o priate a n d t he st u d y desi g ns i ncl u de d a p pr o priate c o ntr ols, 
a n d use d a p pr o priate/ vali date d a nal ytical met h o ds. Details of t hese st u dies are i ncl u de d i n 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 7  of 5 4  
 
 
t he re vise d s u b missi o n ( S N 0 0 0 1).  
 
1. 1 0  
Cli nic al St u d y Pl a n  
 
T he pri mar y g o al of t his h u ma n pil ot st u d y is t o tra nslate t he n o v el i ma gi n g a ge nt L S 3 0 1, 
w hic h selecti vel y i de ntifies breast ca ncer cells wit h hi g h acc urac y , t o h u ma n patie nts. 
Di verse a ni mal m o dels of breast ca ncer ha v e bee n i n vesti gate d  t o s ur gi cal re m o val of 
s p o nta ne o us, disse mi nate d, a n d diff use t u m ors, w hic h were detecte d t o si n gle ca ncer cell 
le vel b y L S 3 0 1. We ha ve als o s h o w n t hat t hat t he NI R fl u oresce nce ca n be ca pt ure d 
efficie ntl y usi n g C V G.  
 
T he ma xi m u m r ec o m me n de d starti n g d os e ( M R S D) f or t his cli nical trial w as base d o n t he 
N O A E L fr o m a ni mal st u dies. As s h o w n i n t he G L P t o xic ol o g y d oc u me nt, t he N O A E L 
fr o m t he rat t o xicit y st u d y is 1 0 m g/ k g f or fe males a n d 0. 5 m g/ k g f or male rats, w hic h is 
e qui vale nt t o h u ma n e q ui vale nt d ose ( H E D ) of 1. 6 m g/ k g a n d 0. 0 8 1 m g/ k g, res pecti vel y. 
A p pl yi n g a safet y mar gi n (t y picall y 1 0 -f ol d) t o t he N O A E L gi ves M R S D of 0. 1 6 m g/ k g 
base d o n fe male N O A E L a n d 0. 0 0 8 1 m g/ k g base d o n t he N O A E L i n male rats. T he hi g hest 
n o n-s e ver el y t o xic d ose ( H N S T D)  i n t he rat t o xicit y st u d y was deter mi ne d t o be 1 0 m g/ k g 
( H E D 1. 6 m g/ k g) i n b ot h male a n d fe male rats.  A n ot her a p pr oac h t hat is c o nsi dere d 
a p pr o priate t o deter mi ne a safe starti n g d ose is usi n g 1/ 6 t h of t he H N S T D i n a m ore 
se nsiti ve s pecies. Bas e d o n t he N O A E L a n d H N S T D fr o m t he rat t o xi cit y st u d y, t he 
ma xi m u m rec o m me n de d safe d ose t o test i n t he FI H st u d y was esti mate d t o be bet wee n 
0. 1 6 a n d 0. 2 6 m g/ k g. W hile pre vi o us i n vesti gati o ns wit h L S 3 0 1 i n mice s h o we d t he l o w est 
detecta ble d ose wit h t h e C V G t o be 0. 0 7 5 m g/ k g (e q ui vale nt t o H E D of t hat is 0. 0 0 6 
m g/ k g), a n a ntici pate d o pti mal starti n g d ose of 0. 1 m g/ k g will be use d as t h e starti n g d os e 
i n t he cli nical st u d y. T his d ose is well wit hi n t he esti mate d ra n ge f or t he M R S D esti mate d 
fr o m t he rat t o xicit y st u d y.  
 
We pr o p ose a p hase 1 r olli n g si x d ose- escalati n g desi g n gi ve n  t hat t his is t he first-i n- h u ma n 
use of L S 3 0 1, f oll o we d b y a p h ase 2 trial f or preli mi nar y e v al uati o n of its dia g n ostic 
ca pa bilities  (i ncl u di n g  se nsiti vit y  a n d  s pecificit y).   D uri n g  p hase  1 ,  L S 3 0 1  will  be  
a d mi nistere d i ntra ve n o usl y t o a c o h ort of si x patie nts at a d ose of 0. 0 5 m g/ k g; if fe wer t ha n 
2 D L Ts ar e o bser v e d, L S 3 0 1 will be gi ve n at t he ne xt hi g h d ose le vel of 0. 0 7 5 m g/ k g; if 
fe wer t ha n 2 D L Ts are o bser ve d, L S 3 0 1 will be gi ve n at t he e x p ecte d o pti mal d ose of 0. 1 
m g/ k g .  O nce safet y is c o nfir me d a n d t he o pti mal i ma gi n g d ose is esta blis he d , a n a d diti o nal 
9 patie nts will be e nr olle d i n t he p hase 1 e x pa nsi o n c o h ort t o e val uate f or s afet y. 
 
After p h ase 1 d ata has bee n s u b mitte d t o a n d disc usse d wit h t he F D A, a n a d diti o nal c o h ort 
of 8 8 patie nts will be e nr olle d t o t he p hase 2 p orti o n of t he trial t o assess t he dia g n ostic 
ca pa bilities of L S 3 0 1 f or i de ntificati o n of p ositi ve mar gi ns at s ur ger y.  Base d o n precli nical 
st u dies3 6 ,3 7 ,4 0 ,4 3 ,6 8, t he a ntici pate d o pti mal d ose f or L S 3 0 1 is 0. 1 m g/ k g b o d y wei g ht ( B W). 
T his d ose is a fracti o n of I C G c o n ce ntrati o n c urre ntl y a p pr o ve d f or h u ma n use . L S 3 0 1 will 
be f or m ulate d i n 1 % h u m a n ser u m al b u mi n ( H S A) i n sali ne a n d a d mi nistere d i ntra ve n o usl y 
as a sl o w i nf usi o n o ver 5 -1 0 mi n utes. Base d o n L S 3 0 1 i n 1 % H S A a n d ass u mi n g a 6 0 k g 
a d ult wit h 1. 6 m 2 s urface area a n d usi n g t he c o n v ersi o n fact or k m = 6 0/ 1. 6 = 3 7 k g/ m 2, the 
e q ui vale nt d ose is 1 0 0 µ g/ k g × 3 7 k g/ m 2 = 3. 7 m g/ m2.  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 8  of 5 4  
 
 
 
I n all patie nts, L S 3 0 1 will be a d mi nistere d 4- 2 4 h o urs bef ore l u m pect o m y. L S 3 0 1 u pta ke 
i n pri mar y breast ca ncer or S L N vers us u ni n v ol ve d tiss ue will be vis ualize d wit h t he C V G 
s yste m d uri n g s ur ger y b y a me m b er  ( o p erati n g or n o n- o perati n g) of t h e s ur gical tea m . T he 
NI R li g ht s o urce f or t he C V G is base d o n li g ht -e mitti n g di o des ( L E Ds)  or laser li g ht 
o perati n g at 1 5 -2 0 m W/c m 2 o ut p ut at tiss ue, w hic h ca n deli ver s ufficie nt li g ht t o bi ol o gical 
tiss ues a n d i n d uce fl u oresce nce e missi o n t o meet t he nee ds of t he pla n ne d cli nical st u dies. 
It s h o ul d be n ote d t hat t h e li g ht s o urce is bel o w t h e U S F D A rec o m me n de d li mit f or NI R 
e x p os ure a n d A N SI sta n dar d. T he fl u oresce nce si g nals will be recei ve d b y t he detect or 
p orti o n of o ur de vi ce. G ai n -setti n gs c o ul d be easil y a dj uste d d uri n g o perati o n t o o pti mize 
t he c o ntrast bet wee n hi g h fl u oresce n ce areas (t u m ors) a n d l o w fl u ores ce n ce areas ( n or mal 
tiss ues). Real-ti me fl u oresce nce i ma g e will be dis pla ye d i n t he g o g gle e ye piece as w ell as 
o n a sec o n dar y m o nit or t o facilitate vie wi n g b y ot her me m bers of t he s ur gi cal tea m i n t he 
O R . I ma g es fr o m t he C V G or S P Y will n ot be use d t o alter t he s ur gical pla n i n a n y wa y. 
 
Fl u oresce nce i n t he e xcis e d tiss ue at t he mar gi ns will be deter mi ne d b y q u a ntitati ve near -
i nfrare d fl u ores ce nce micr osc o p y. F or eac h p atie nt, at least 4 s peci me ns fr o m e xcise d 
tiss ue  will  be  a nal yze d,  i ncl u di n g  s us pecte d  t u m or  a n d  s us pecte d  h ealt h y  tiss ue. De -
i de ntifie d data will be st ore d o n a c o m p uter. St ore d i nf or mati o n will o nl y ha ve c o de d 
n u m bers a n d n ot t he na m es of t he patie nts t o e ns ure patie nce pri vac y. 
   
1. 1 1  
R ati o n ale 
 
O ur  precli nical  data  ha ve  de m o nstrate d  t he  feasi bilit y  of  fl u oresce nce -g ui de d  t u m or  
resecti o n b y o ur C V G  wit h   L S 3 0 1 i n a ni mal m o dels. I n t his st u d y, we will  c o n d uct  
i ntra o perati ve i ma gi n g pr oce d ures t hat ha v e mi ni mal i nterfere nce wit h o n g oi n g s ur ger y. 
T he u n del yi n g h y p ot h esis is t hat t he acc urat e det ecti o n of all ca ncer cells hi g hli g hte d b y 
L S 3 0 1  d uri n g  s ur ger y  will  re d uce  t he  n u m ber  of  breast  ca ncer  p atie nts  wit h  mar gi n  
p ositi vit y t o less t ha n 5 %, c o m pare d t o t he c urre nt s ur gical para di g m of greater t ha n 2 0 % . 
T he pil ot st u d y will o btai n critical data re q uire d t o a d dress t he lar g er q uesti o n of s ur gical 
mar gi n assess me nt i n a f ull p hase I cli nical trial.  
 
1. 1 2  
P ote nti al C o ntri b uti o n  
 
T he a bilit y t o i de ntif y t u m ors i n viv o w o ul d be a hi g hl y usef ul cli nical t o ol f or o nc ol o gic 
s ur ge o ns.  W he n  utilize d  i n  t u m or  resecti o n,  i ncl u di n g  S L N  bi o ps y,  t he  pr o p ose d  
pr oce d ur es ca n rea dil y detect t u m or mar gi ns, s mall n o d ules, resi d ual t u m ors, a n d p ositi ve 
S L Ns, t here b y re d uci n g t he i nci de nce of i nc o m plete resecti o n, t he size of h ealt h y tiss ues 
resect e d, a n d t h e nee d f or re visi o n s ur g eries. M ore o ver, t h e pr o p ose d tec h n ol o g y ca n als o 
offer hi g h s p atial res ol uti o n i n real -ti me wit h o ut t he use of ra di oacti ve tracers.  I n a d diti o n 
t o i m pr o vi n g healt hcar e, t he a p pr o ac h  ca n p ote ntiall y s h orte n O R ti me t o f urt her re d u ce 
c ost of s ur ger y. T heref or e, t he L S 3 0 1 -me dia te d fl u oresce n ce i ma gi n g of t u m ors has hi g h 
p ote ntial i m pact o n breast ca ncer ma na g e me nt.  
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 1 9  of 5 4  
 
 
2. 0  
O B J E C TI V E S  
 
2. 1  
Pri m ar y O bjecti ves  
 
P hase 1 :  to deter mi ne t h e saf et y a n d o pti mal i ma gi n g d ose of L S 3 0 1 i nj ecte d i n breast 
ca ncer patie nts.  
 
P hase 2 : t o deter mi ne t he a bilit y of t his n o vel fl u oresce n ce i ma gi n g a ge nt t o pre dict t he 
prese nce of p ositi ve mar gi ns ar o u n d partial mastect o m y s peci me ns a n d p ositi ve S L Ns 
d uri n g s ur gical t her a p y f or breast ca ncer. 
 
3. 0  
P A TI E N T S E L E C TI O N  
 
3. 1  
I ncl usi o n Criteri a 
 
1.  N e wl y  dia g n ose d  Sta ge  I -II  breast  ca ncer  pati e nts  u n der g oi n g  breast-c o nser vi n g 
t hera p y a n d S L N  bi o ps y. 
2.  N e gati ve n o dal b asi n cli nical e xa m.  
3.  At least  1 8 years of a ge.   
4.  A ble t o u n dersta n d a n d willi n g t o si g n a writte n i nf or me d c o ns e nt d oc u me nt. 
 
3. 2  
E xcl usi o n Criteri a  
 
1.  C o ntrai n dicati o ns f or s ur ger y.  
2.  Recei vi n g a n y i n vesti gati o nal a ge nts. 
3.  H ist or y of aller gic reacti o ns attri b ute d t o I C G or ot her a g e nts use d i n t he st u d y, i ncl u de 
k n o w n i o di de or seaf o o d aller g y . We d o n ot e x pect ma n y of t hese a d vers e reacti o ns 
wit h L S 3 0 1 beca use it is n ot ra di oacti ve a n d d o es n ot p ossess i o di nate d c o u nteri o ns. 
4.  Prese nce of u n derl yi n g l u n g disease.   
5.  Pre g na nt.   Fe m ale p atie nts of c hil d beari n g p ote ntial m ust ha ve a n e gati ve ser u m or 
uri ne pre g n a nc y test n o m ore t ha n 7 da ys bef ore start of partici pati o n. 
6.  Breastfee di n g. Patie nts w h o are breastfee di n g are e xcl u de d fr o m t his st u d y beca use 
t here is a n u n k n o w n b ut p ote ntial ris k f or a d v erse e ve nts i n n ursi n g i nfa nts sec o n d ar y 
t o treat me nt of t he m ot her wit h L S 3 0 1.  
 
3. 3  
I ncl usi o n of W o m e n a n d Mi n orities  
 
Breast ca n cer is r are i n m e n; t heref ore, it is li kel y t hat o nl y w o me n will e nr oll i n t his trial , 
alt h o u g h me n are eli gi ble.  Pe o ple of all races a n d et h nicities ma y e nr oll.  
 
4. 0  
R E GI S T R A TI O N P R O C E D U R E S  
 
P atie nts m ust n ot st art a n y pr ot oc ol i nter v e nti o n pri or t o re gistr ati o n t hr o u g h t he Site m a n 
C a ncer Ce nt er  f or p h ase 1 A a n d U T S o ut h wester n Me dic al Ce nter V el os d at a b ase f or p h ase 
1 B a n d p h ase 2 . 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 0  of 5 4  
 
 
T he f oll o wi n g ste ps m ust be ta ke n bef ore re gisteri n g patie nts t o t his st u d y: 
 
1.  C o nfir mati o n of patie nt eli gi bilit y  
2.  Re gistrati o n of patie nt i n t he Site ma n Ca ncer Ce nter O n C ore data b ase  f or p hase 1 A a n d 
U T S o ut h wester n Me dical Ce nter Vel os dat a base f or p hase 1 B a n d  ph ase 2.  
3.  Assi g n me nt of u ni q ue patie nt n u m ber ( U P N) 
 
4. 1  
C o nfir m ati o n of P atie nt Eli gi bilit y  
 
C o nfir m patie nt eli gi bilit y b y c ollecti n g t he i nf or mati o n liste d bel o w:  
 
1.  Re gisteri n g M D’s na me 
2.  Patie nt’s race, s e x, a n d D O B  
3.  T hree letters ( or t w o letters a n d a das h) f or t he p atie nt’s i nitials  
4.  C o p y of si g ne d c o nse nt f or m 
5.  C o m plete d eli gi bilit y c hec klist, si g ne d a n d date d b y a me m ber of t h e st u d y tea m 
6.  C o p y of a p pr o priate s o urce d oc u me nt ati o n c o nfir mi n g patie nt eli gi bilit y 
 
4. 2  
P atie nt Re gist r ati o n i n t he Site m a n C a ncer Ce nter O n C ore D at a b ase f or 
p h ase 1 A U T S o ut h wester n Me dic al Ce nter V el os D at a b ase f or p h ase 1 B 
a n d p h ase 2. 
 
All patie nts m ust be re gistere d t hr o u g h t he Site ma n Ca ncer Ce nter O n C ore data b ase  f or 
p hase 1 A U T S o ut h wester n Me dical Ce nter Vel os data base f or p has e 1 B a n d  phase 2 . 
 
4. 3  
Assi g n m e nt of U P N  
 
Eac h p atie nt will be i de ntifie d wit h a u ni q ue p atie nt n u m ber ( U P N) f or t his st u d y.  All dat a 
will be rec or d e d wit h t his i de ntificati o n n u m ber o n t he a p pr o priat e C R Fs. 
 
4. 4  
B aseli ne D at a C ollecti o n 
1.  Patie nt de m o gra p hic i nf or mati o n: a ge, se x, race, et h nicit y 
2.  Patie nt hei g ht, wei g ht, a n d b o d y mass i n de x ( B MI) 
3.  Patie nt me dical hist or y a n d c o nc o mita nt me dicati o ns  
 
5. 0  
S T U D Y P R O C E D U R E S  
 
5. 1  
L S 3 0 1 Pre p ar ati o n  
 
L S 3 0 1 will be pre p are d u n der c G M P pr ot oc ol a n d s u p plie d i n a n a m b er -c ol ore d vial as a 
l y o p hilize d pr o d uct i n 1 % H S A/ P B S a q ue o us s ol uti o n. Eac h vial will c o ntai n 2 m g L S 3 0 1. 
Bef ore a d mi nistrati o n, 1 0 m L of P B S will be a d de d t o t he l y o p hilize d p o w der a n d mi xe d 
b y s ha ki n g f or 1 mi n ute. T he a q ue o us f or m ulati o n is sta ble at r o o m te m perat ure f or 4 h o urs 
or i n t he refri ger at or ( 2- 8° C) f or 2 4 h o urs. T he a ge nt will be a d mi nistere d 4 -2 4 h o urs b ef ore 
s ur ger y.   
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 1  of 5 4  
 
 
5. 2  
Pr oce d ures  
 
5. 2. 1  P h ase 1  
 
A r olli n g si x desi g n (refer t o Secti o n 8. 1) will be use d t o e nr oll patie nts w h o are 
bei n g treate d f or p at h ol o gicall y pr o ve n breast ca ncer pri or t o defi niti ve s ur gical 
i nter ve nti o ns i n t he p hase 1 A d ose escalati n g p orti o n of t he st u d y, d uri n g w hic h 
L S 3 0 1 will be a d mi nistere d i ntra ve n o usl y at o ne of t hree d oses (see ta ble bel o w) .  
Patie nts  will  be  o bser ve d f or  d ose-li miti n g  t o xicities  ( D L T) (refer  t o  Secti o n  
5. 2. 1. 3). 
 
L S 3 0 1 D ose Le vel  
L S 3 0 1 D ose  
Le vel 1 (starti n g d ose)  
0. 0 5 m g/ k g  
Le vel 2  
0. 0 7 5 m g/ k g  
Le vel 3  
0. 1 m g/ k g  
 
T he patie nt will u n der g o s ur ger y 4- 2 4 h o urs after a d mi nistrati o n of L S 3 0 1 usi n g 
c urre nt b est pr actices.  Breast tiss ue  a n d a xillar y L Ns  will be i ma ge d n o n-i n vasi vel y 
pri or t o start of s ur ger y a n d e xcise d breast tiss ue a n d S L Ns will be e xa mi ne d e x 
vi v o f or t h e pr ese nce of L S 3 0 1 fl u ores ce n ce usi n g o ur Ca n cer Visi o n G o g gles 
( C V G) t o deter mi ne if L S 3 0 1 acc u m ulate d i n t he breast ca n cer. We will q ua ntif y 
fl u oresce n ce i nte nsit y i n t he ca ncer t o esta blis h t he feasi bilit y of o bser vi n g L S 3 0 1 
fl u oresce n ce wit h t he i m a gi n g s yste m. 
 
T he d ose us e d i n c o h ort 1 is  t he l o west d ose ( 0. 0 5 m g/ k g) a n d was use d e xcl usi vel y 
f or safet y assess me nt. T h e i n vesti gat ors di d n ot e x pect t o d etect L S 3 0 1- ass ociate d 
t u m or tiss ue  fl u oresce nce bef ore  i ncisi o n  at  t h is l o west  d ose  le vel. T heref ore, 
c o h ort  1  patie nts  will  n ot  be  i ma ge d  n o n-i n v asi vel y t o  assess  t u m or tiss ue 
fl u oresce n ce bef ore i ncisi o n. N o n -i n vasi ve t u m or fl u oresce n ce e xa mi nati o n will be 
i nitiate d fr o m c o h ort 2 o n war ds, w here hi g her d oses will be use d. 
 
I n v esti gat ors d o n ot e x pect t he 1st d ose ( 0. 0 5 m g/ k g) or 2 n d d ose ( 0. 0 7 5 m g/ k g) t o 
lea d t o a n y detecta ble fl u oresce n ce n o n-i n vasi v el y i n t he a xilla bef ore se nti nel 
l y m p h n o de e xcisi o n d u e t o t he l o w c o nce ntrati o n of e x p ecte d L S 3 0 1 re t e nti o n i n 
l y m p h  n o des.  T heref ore,  l y m p h  n o de  fl u ores ce nce  will  n ot  be  assesse d  n o n-
i n vasi vel y bef or e i ncisi o n f or c o h orts 1 a n d 2. I n vesti gat ors will pla n t o i ma ge 
l y m p h n o de re gi o ns n o n-i n vasi vel y starti n g wit h c o h ort 3 d ue t o t he hi g her i njecte d 
d ose ( 0. 1 m g/ k g). 
 
After t he o pti mal d ose h as bee n deter mi ne d, a n a d diti o nal 9 patie nts will be e nr olle d 
i n t he p hase 1 e x pa nsi o n c o h ort t o e val uate f or safet y. T his e x pa nsi o n c o h ort is  
c o nsi dere d p h ase 1 B a n d will be c o n d ucte d at U T S o ut h wester n Me dical C e nter.  
 
 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 2  of 5 4  
 
 
5. 2. 1. 1  D ose Esc al ati o n Sc he m a  
 
L S 3 0 1 D ose Le vel  
L S 3 0 1 D ose  
Le vel 1 (starti n g d ose)  
0. 0 5 m g/ k g  
Le vel 2  
0 .0 7 5 m g/ k g  
Le vel 3  
0. 1 m g/ k g  
 
D ose  escalati o n  ma y  o cc ur  as  s o o n  as  t he  last  patie nt  i n  a  c o h ort  has  
c o m plete d her o n e- h o ur p ost -i njecti o n t o xicit y m o nit ori n g. 
 
5. 2. 1. 2  Defi niti o n of O pti m al D ose  
 
T he o pti mal d ose will be t he d ose at w hic h fe wer t ha n 2 D L Ts are o bser v e d  
a n d  o pti mal  i ma ge  q ualit y  is  o bser ve d.   It  n ee d  n ot  be  t he  ma xi m u m  
t olerate d d ose if o pti mal i ma ge q ualit y is o bser v e d at a l o w er d os e t ha n t h e 
M T D.  
 
5. 2. 1. 3  Defi niti o n of D ose -Li miti n g T o xicities  
 
D L T is defi ne d as a n y gra de 2 or a b o v e t o xicit y t hat occ urs d uri n g t he 
h o ur after i njecti o n t hat is c o nsi dere d p ossi bl y, pr o ba bl y, or defi nitel y 
relate d t o L S 3 0 1. 
 
5. 2. 1. 4  D ose Esc al ati o n Criteri a  
 
T he m olec ular pr o b e L S 3 0 1 is a n i ma gi n g a ge nt bi n di n g t o breast ca ncer 
cells a n d ot her mali g na nt cells a n d will be vis ualize d t hr o u g h b y C V G 
w hic h is a n o n -si g nifica nt ris k de vice. Si milar t o t he F D A-a p pr o v e d a ge nt 
I C G, w e d o n ot e x pect a n y seri o us a d vers e e v e nts ca use d b y t he a ge nt.  
 
A r olli n g si x desi g n will be use d f or t he p h ase 1 p orti o n.  I n t he r olli n g si x 
desi g n, 2 t o 6 patie nts ca n be e nr olle d t o a d ose eac h ti me.  T he d ose de -
escalati o n o cc urs w he n t w o or m ore D L Ts occ ur at a d os e le v el, w hile d os e 
escalati o n occ urs wit h 3/ 3/, 4/ 4, 5/ 5, 5/ 6, or 6/ 6 patie nts are e v al uate d 
wit h o ut D L T.  ( Refer t o Secti o n 8. 1 f or f urt her details.)   
 
5. 2. 2  P h ase 2  
 
After  data fr o m  t he p hase 1 p orti o n of t he st u d y have bee n s u b mitte d t o a n d 
re vie we d b y t he F D A, a n d if a nal ysis of t he data fr o m p hase 1 c o nfir ms t he safet y 
of a d mi nisteri n g L S 3 0 1 i n h u ma ns  a n d esta blis hes t he o pti mal i ma gi n g d ose , 8 8 
patie nts will be e nr olle d i n t he p hase 2 p orti o n of t he st u d y  aft er a n y n ecessar y 
pr ot oc ol re visi o ns ha ve b ee n ma de b ase d o n p has e 1 data a n d F D A a p pr o val.   
 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 3  of 5 4  
 
 
It  is  e x p ecte d  t hat  t he  st u d y  pr oce d ures  f or  p h ase  2  will  be  t he  s a me  as  t he  
pr oce d ur es i n p has e 1.  T he o pti mal i ma gi n g d ose of L S 3 0 1 deter mi ne d i n p hase 1 
will be a d mi nistere d i n p hase 2. 
 
T he breast tiss ue  a n d a xillar y L Ns will be i ma ge d n o n -i n vasi vel y usi n g t he C V G 
a n d w or n b y a n o n -o p erati n g me m ber of t he s ur gical tea m. After re m o v al of t he 
breast ca ncer a n d S L Ns usi n g c urre nt best practice, a me m ber of t he s ur gical tea m  
w h o is n ot c o n d ucti n g t he s ur g er y will wear t h e C V G t o deter mi ne if L S 3 0 1 
fl u oresce n ce is pr ese nt at t he mar gi n a n d i n S L Ns, usi n g a gl o bal t hres h ol d of 2 0 % 
si g nal a b o ve bac k gr o u n d or a d y na mic statistical t hres h ol d t o i n dicate a p ositi ve 
mar gi n.  T he acc ur ac y of deter mi ni n g mar gi n p ositi vit y will be esta blis he d b y 
c o m pari n g t he C V G ass ess me nt t o hist o pat h ol o g y. 
 
5. 2. 3  A d mi nistr ati o n of L S 3 0 1 
 
T he  patie nt  will  u n der g o  i ntra ve n o us i njecti o n  of  L S 3 0 1  4-2 4  h o urs  pri or  t o  
s ur ger y.  A st u d y n urse or ot her q ualifie d me m ber of t he st u d y tea m will i nject t he 
L S 3 0 1 via I V o ver 5- 1 0 mi n utes i n t he office or pre -o p r o o m  (as a p plica ble) a n d 
will m o nit or t he patie nt as descri be d i n Secti o n 5. 4 bel o w. A d mi nistrati o n of L S 3 0 1 
will n ot i nterfer e wit h t he sta n dar d of care, i ncl u di n g all sta n dar d of care f or l y m p h 
n o de assess me nt usi n g 9 9 mTc Til ma n oce pt  i njecti o n o n t he da y of or d a y bef ore 
s ur ger y. 
 
5. 2. 4  I ntr a o per ati ve R e vie w 
 
T he o perati n g s ur g e o n will c o n d uct breast ca ncer s ur ger y as us ual wit h o ut usi n g a 
de vice t o vis ualize L S 3 0 1 fl u oresce nce. T o pre v e nt bias i n data ac q uisiti o n, a n o n-
o perati n g me m ber of t he st u d y tea m  will wear t he C V G  bef ore t he start of s ur ger y 
t o e xa mi ne t he br east tiss ue a n d a xillar y L Ns  n o n-i n vasi vel y a n d at t he c o m pleti o n 
of t he s ur ger y t o e x a mi ne t he e xcise d tiss ue , S L Ns  a n d s ur gical ca vit y . Fi n di n gs 
wit h t he st u d y will be rec or de d b ut will n ot c ha n ge t he sta n dar d of care.  T he 
o perati n g s ur g e o n will re mai n bli n de d t o t he fl u oresce n ce i ma g es t hr o u g h o ut t he 
o perati o n. T he e x a mi ni n g s ur gical tea m me m ber will wear t he C V G a n d i ma ge all 
si x a nat o mical as pects (s u peri or, i nferi or, a nteri or, p osteri or, me dial a n d lateral) of 
t he s ur gical s peci me n as well as t he s ur gical ca vit y f or L S 3 0 1 fl u oresce n ce. T he 
S P Y s yste m will be use d f or c o nfir mat or y fl u oresce nce i ma gi n g a n d c o m pare d wit h 
C V G detecti o n of L S 3 0 1 fl u oresce n ce, d uri n g p hase 2 of t he st u d y. 
 
Detecti o n of L S 3 0 1 fl u oresce n ce at t he c ut e d ge of t he e xcise d s peci me n will 
deter mi ne mar gi n p ositi vit y  a n d prese n ce of fl u oresce n ce i n t h e S L Ns i n dicate 
prese nce of metastatic ca ncer cells . Gi ve n t hat C V G ca n detect L S 3 0 1 fl u oresce nce 
i n t u m ors se veral milli meters dee p, i n vesti gat ors will use pat h ol o g y data t o i nitially 
trai n t he s yste m b y c orrelat i n g fl u ores ce nce i nte nsit y t o bac k gr o u n d rati o wit h 
pat h ol o gist’s mar gi n ass ess me nt . T he i nf or mati o n will t he n be use d i n p h ase 2 t o 
pre dict mar gi n p ositi vit y i n t he O R. T he d etecti o n of fl u oresce n ce o n e xcise d tiss ue 
wit h o ut c o rrelati ve fl u oresce nce i n t he t u m or ca vit y after l u m pect o m y will pre dict 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 4  of 5 4  
 
 
a ne gati ve mar gi n f or i n v asi ve d uctal carci n o ma. F or D CI S, laser p o wer m o d ulati o n 
wit hi n t he a p pr o ve d li mit or d ual e xcitati o n or e missi o n met h o d will be use d t o 
esti mate t he d e pt h of fl u oresce n ce s o urce. I n ge n eral, a  si g nal t o bac k gr o u n d rati o 
of 1. 2 ( 2 0 % hi g her i nte nsit y t ha n bac k gr o u n d) will be use d as a gl o bal t hres h ol d t o 
deter mi ne mar gi n a n d S L N p ositi vit y  stat us. I n a d diti o n, a d y na mic statistical 
t hres h ol di n g a p pr oac h will be use d t o deli neate t he b o u n dar y of tiss ue fl u oresce nce . 
A n i nte nsit y-base d  heat c ol or ma p will be use d f or false c ol or re prese ntati o n of t he 
fl u oresce n ce t o g ui de t he e xa mi ni n g s ur ge o n. Areas wit h hi g h L S 3 0 1 acc u m ulati o n, 
will be r e pres e nte d as r e d -h ot c o m pare d t o gree n a n d bl ue f or s urr o u n di n g tiss ue. 
A c o nti n u u m of re d fl u oresce n ce will i n dicate t hat t he e ntire e xcise d tiss ue is 
ca ncer.  Fl u oresce nce gra d ie nt usi n g t he heat c ol or ma p s h o wi n g re d t o bl ue fr o m 
t he e d ge of t u m or tiss ue will i n dicate a p ositi ve mar gi n. W he n t he fl u oresce nce 
e ma nati n g fr o m t u m ors a wa y fr o m t he mar gi n ( > 2 m m), haz y bl ue fl u oresce n ce 
will be o bser ve d at t he s ur gical mar gi n  a n d will be classifie d as cl ear mar gi n .   
 
If t his si m ple a p pr oac h fails t o acc uratel y i de ntif y p ositi ve or clea n m ar gi ns, we 
will  use  a  d ual  wa vele n gt h  e xcitati o n  f or  t he  de pt h -pr ofili n g  of  fl u oresce nce  
e missi o n. It  is  well  k n o w n  t hat  li g ht  tra vels  t hr o u g h  tiss ue  differe ntl y  acr oss  
wa vele n gt hs d ue t o wa vele n gt h -de pe n de nt o ptical pr o perties. W he n a fl u or o p h ore 
is e xcite d  usi n g  t w o  wa vele n gt hs,  t he  i nte nsit y  fall off  wit h  de pt h  of  eac h  
wa vele n gt h  will  be r elate d  t o  t he  de pt h  of  t he  fl u or o p h ore.  B eca us e  b ot h  
wa vele n gt hs  ar e  e xciti n g  t he  sa me  fl u or o p h ore  c o nce n trati o n,  t he  rati o  of  t he  
fl u oresce n ce  g e nerate d  yiel ds  t he  de pt h  of  t he  fl u or o p h ore  irres p ecti ve  of  
c o nce ntr ati o n  acc u m ulat e d i n t he t u m or tiss ue . The de pt h of fl u oresce n ce i n t he 
resect e d sa m ple will be esti mate d usi n g t w o s pectrall y se parat e d NI R w a vel e n gt hs 
t o e xcite L S 3 0 1 fl u oresce nce. T h e rati o of t he e mitte d i nte nsities i ma ge d f or b ot h 
e xcitati o n wa vel e n gt hs will be cl oser t o o n e w h e n t he t u m or tiss ue is at t he c ut 
s urface.  At  hi g her  d e pt hs  t he  rati o  will  de viat e  a w a y  fr o m  1  as  t he  s h orter  
wa vele n gt h  e xcitati o n is atte n uate d m ore t ha n l o n ger e x citati o n. A de pt h ma p of 
tiss ue fl u oresce nce will be create d a n d c o m pare d t o hist o pat h ol o g y f or deter mi ni n g 
t he acc ur ac y of pr e dicti n g p ositi ve or clear mar gi n stat us  a n d S L N metastatic stat us . 
T he d ual- w a vele n gt h m et h o d will i nitiall y be i m ple me nte d as a sta n d al o ne p orta ble 
cl ose d b o x c o nfi g urati o n w hic h will t he n be mi niat urize d a n d i m ple me nte d i n t he 
C V G.  
  
T he s ur gicall y re m o ve d t u m or mass a n d S L Ns will be pat h ol o gicall y e xa mi ne d 
u n der sta n d ar d pr ot oc ols. At least 4 s peci me ns fr o m t he e x cise d tiss ue of a patie nt, 
t w o s us pecte d t u m or a n d t w o s us pecte d healt h y tiss ue per patie nt will be a n al yze d. 
 
T he t u m or s peci me ns ge nerate d after brea dl oafi n g b y t he s ur gi cal pat h ol o gist will 
be i n di vi d uall y i ma ge d usi n g t he C V G a n d t he d ual -wa vele n gt h cl ose d b o x i ma gi n g 
s yste m bef ore fi xati o n a n d paraffi n e m be d di n g t o rec or d t he fl u oresce nce i nte nsit y 
l ocalizati o n a n d de pt h pr ofiles. Pat h ol o gist will pr o vi de tiss ue sa m ples t hat will be 
secti o ne d a n d m o u nte d o n sli des. T he secti o ne d tiss ue sli des will be i m a ge d f or 
L S 3 0 1 fl u oresce nce usi n g hi g h -res ol uti o n c o nf ocal micr osc o pes. T h ese will be 
c o m pare d t o sta n d ar d H e m at o x yli n & E osi n stai ni n g of t u m or tiss ue t o verif y 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 5  of 5 4  
 
 
L S 3 0 1  s pecificit y  f or  ca ncer  cells.  A d diti o nal  i m m u n o hist oc he mistr y  will  be 
perf or me d  o n  t hes e  tiss ue  secti o ns  t o  v erif y  or  i de ntif y   L S 3 0 1-ass ociate d 
bi o mar kers.   
  
5. 3  
E v al u a bilit y 
 
All  patie nts  w h o  recei ve  a n  L S 3 0 1  i njecti o n  are  e val ua bl e  f or  t o xicit y.   Patie nts  are  
e val uate d fr o m ti me of i njecti o n t o o ne- h o ur p ost-i njecti o n. 
 
Patie nts e nr olle d i n p h ase 1  are  e val u a ble f or D L T  assess me nt  d uri n g  t he h o ur  aft er 
i njecti o n.  Patie nts i n p hase 2 will n ot be assesse d f or D L T. 
 
5. 4  
S afet y E v al u ati o n  
 
T he safet y of t he L S 3 0 1 will be assesse d b y cl osel y m o nit ori n g t he patie nts ’ vital si g ns a n d 
la b orat or y tests as des cri be d bel o w. 
 
5. 4. 1  Vit al Si g ns  
 
All vital si g ns will be r ec or de d o n t h e case re p ort f or m. Vital si g ns ma y b e o btai ne d 
wit h t he s u bject i n t he s u pi ne or u pri g ht p ositi o n. Care will be ta ke n t o o btai n 
s u bse q ue nt rec or di n gs wit h t he s u bject i n t he sa me p ositi o n (s u pi ne or u pri g ht). 
Alt h o u g h aller gic or ot her i m me diate a d vers e reacti o ns are n ot a ntici pate d, s u bjects 
will be m o nit ore d f or at l east 6 0 mi n utes  p ost i njecti o n i n a n area w here e m er ge nc y 
e q ui p me nt is a vaila ble. Vital si g ns will be o btai ne d pre -i njecti o n ( wit hi n 3 0 mi n utes 
pri or t o i njecti o n of L S 3 0 1), wit hi n 3 0 mi n utes  p ost i njecti o n, at 6 0 mi n utes  p ost 
i njecti o n, a n d pri or t o s ur ger y. Vital si g ns will i ncl u de t he f oll o wi n g: heart rate, 
s yst olic  bl o o d  press ure,  diast olic  bl o o d  press ure,  res pirat or y  rate,  a n d  b o d y  
te m perat ur e. T he f oll o wi n g c ha n ges fr o m baseli ne will be c o nsi dere d n ote w ort h y:   
 
•  Heart r ate  > 2 0 b eats per mi n ute 
•  S yst olic bl o o d press ure > 2 0 m m H g 
•  Diast olic bl o o d press ure > 1 0 m m H g 
 
N ote w ort h y c ha n g es will be d oc u me nte d o n t he C R F, a n d t he y will be mar ke d as 
cli nicall y si g nifica nt or n ot. If cli nicall y si g nifica nt, t he pri nci pal i n vesti gat or will 
assess t he ca us alit y of t he c ha n g e t o t he i njecti o n of L S 3 0 1. Cli nicall y si g nifica nt 
c ha n ges i n vital si g ns will be f oll o we d u p h o url y u ntil t he y ret ur n t o bas eli ne or 
n or mal le vels, or u ntil f ol l o w-u p is n o l o n ger warra nte d. If a cli nicall y si g nifica nt 
c ha n ge of a vital si g n is n ote d, it will be re p orte d o n t he a d vers e e ve nt l o g.  
 
5. 4. 2  Cli nic al L a b or at or y Testi n g 
 
La b orat or y  tests  will  c o nsist  of  t he  f oll o wi n g:  sta n dar d  C B C,  c o m pre h e nsi ve  
meta b olic pa nel a n d uri nal ysis o btai ne d at t he f oll o wi n g ti me p oi nts:  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 6  of 5 4  
 
 
•  Baseli ne: wit hi n 3 0 mi n utes pri or t o i njecti o n of L S 3 0 1 
•  6 0 mi n utes after i njecti o n of L S 3 0 1 
•  A p pr o xi matel y o ne h o ur p ri or t o s ur ger y 
 
A p pr o xi matel y 1 4 ml of bl o o d a n d a uri ne sa m ple will be c ollecte d at eac h ti me 
p oi nt.  F or patie nts e nr olle d i n p hase 1 B a n d p has e 2 of t he st u d y, a  p orti o n of t he 
bl o o d sa m ples fr o m eac h ti me p oi nt will be use d t o i de ntif y t he pre d o mi na nt bl o o d 
pr otei n(s) t hat bi n ds L S 3 0 1 a n d me diates t he bi oa vaila bilit y of t he a ge nt (refer t o 
Secti o n 5. 6).  P hase 1 B refers t o p has e 1 st u d y after c o m pleti n g dr u g safet y a n d 
selecti n g o pti mal d ose f or testi n g at 2 a n d 4 h o urs fr o m i njecti o n t o s ur ger y. T he 
c urre nt pr oject is t hat if 3 patie nts at eac h of t he 3 d ose le vel d o n ot e x p erie nce 
a d verse e ve nts, w e will nee d o nl y 9 patie nts f or t he dr u g safet y st u d y ( p h ase 1 A) 
an d a d diti o nal 9 patie nts  at t he selecte d d os e f or testi n g at 2, 4, a n d > 1 2 h o urs ( 3 
patie nts f or eac h ti me p oi nt) fr o m i njecti o n t o s ur ger y. 
 
T he res ults of sta n dar d of care la b orat or y t esti n g o btai ne d u p t o 3 0 d a ys p ost 
i njecti o n will be re vie we d. A nal ysis of t his will ta ke i nt o acc o u nt t hat t he s u bject 
has u n der g o n e s ur gical re m o val of t u m or a n d n e oa dj u va nt treat me nt i n cl u di n g 
h or m o nal t hera p y or c he m ot hera p y.  
  
T he f oll o wi n g c ha n ges fr o m baseli ne cli nical la b orat or y val ues are c o nsi d ere d t o 
be n ote w ort h y a n d re q uire assess me nt as t o cli nical si g nifica nce w he n t h e y fall 
o utsi de of n or mal li mits. Cli nicall y si g nifica nt c ha n ges i n la b orat or y val ues s h o ul d 
be f oll o we d u p dail y u ntil t he y ret ur n t o baseli ne or n or mal le vels, or u ntil f oll o w -
u p is n o l o n ger warra nte d. La b orat or y val ues t hat are a b n or mal at baseli ne b ut m o ve 
i nt o n or mal ra n ge will n ot be c o nsi dere d cli nicall y si g nifica nt.  If a cli nicall y 
si g nifica nt c ha n ge of a la b orat or y val ue is n ote d, it will be re p orte d o n t he A E  l o g.   
 
A n al yte  
C h a n ge fr o m b aseli ne  
He m o gl o bi n  
> 2 g/ d L  
W B Cs  
> 1 K/ m m 3  
Ne utr o p hils  
> 1 0 %  
L y m p h oc ytes  
> 1 0 %  
Platelets  
> 5 0 K/ m m 3  
Creati ni ne  
> 0. 7 5 m g/ d L  
B U N  
> 2 0 m g/ d L  
Calci u m  
> 1 m g/ d L  
S o di u m  
> 5 m m ol// L  
P otassi u m  
> 0. 5 m m ol/ L  
C O 2  
> 4 m m ol/ L  
A L T ( S G P T)  
> 1 5 0 I U/ L  
A S T ( S G O T)  
> 1 0 0 I U/ L  
Al kali ne P h os p hatase  
> 1 5 0 I U/ L  
T otal Bilir u bi n  
> 0. 5 m g/ d L  
Al b u mi n  
> 1 g/ d L  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 7  of 5 4  
 
 
 
C ha n ges i n pre a n d p ost i njecti o n uri nal ysis will be n ote d b ut d u e t o varia bilit y will 
n ot be use d t o deter mi ne cli nical si g nifica nce f or c ha n ges d ue t o t he i njecti o n of 
L S 3 0 1.     
 
 
5. 4. 3  Electr oc a r di o gr a ms ( E C Gs)  
 
A sta n dar d 1 2 -lea d E C G will be o btai ne d o n all s u bjects at bas eli ne ( wit hi n 3 0 
mi n utes pri or t o i njecti o n of L S 3 0 1), 5 -1 0 mi n utes  p ost i njecti o n, a n d pri or t o 
disc har ge at 6 0 mi n utes.   
 
T he f oll o wi n g ta ble lists criteria f or n or mal li mits a n d cli nicall y n ota ble li mits f or 
E C Gs i n a d ults.  
 
 
N or m al Li mits ( msec)  
N ot a ble Li mits ( msec)  
E C G V ari a bles  L o w  
Hi g h  
L o w  
Hi g h  
P R i nter val  
1 2 0  
2 0 0  
< 1 2 0  
> 2 0 0  
Q R S i nter val  
5 0  
1 0 0  
< 5 0  
> 1 0 0  
R R i nter val  
6 0 0  
1 0 0 0  
< 6 0 0  
> 1 0 0 0  
Q T i nter val  
N o l o wer li mit  
≥ 4 6 0  
N o l o wer li mit  
≥ 4 6 0  
 
5. 4. 4  I de ntific ati o n of Bl o o d h alf-life of L S 3 0 1 
 
L S 3 0 1 half- life will be d eter mi ne d usi n g 5 0 0 µ L of bl o o d c ollecte d at t he sa me 
ti me as t he C B C a n d C M P at baseli ne, 6 0 mi n utes p ost-i njecti o n, a n d o ne h o ur 
bef ore s ur ger y . T h ese bl o o d sa m ples f or researc h use will be dra w n i nt o pi n k t o p 
li g ht- pr otecte d t u bes, br o u g ht t o t he Ac hilef u la b f or a nal ysis, t o i de ntif y t he 
c o nce ntr ati o n of L S 3 0 1 a n d deter mi ne of t he h alf- life of t he a g e nt.   
 
5. 5  
Ris ks a n d A d verse E ve nt M o nit ori n g  
 
A p ote ntial ris k s pecific t o t his e x peri me nt pr o ce d ure is t he i njecti o n of L S 3 0 1. T he 
i ma gi n g a ge nt is n ot k n o w n t o ca use a n y s eri o us a d vers e eve nts i n precli nical m o dels. 
H o we ver, it is p ossi ble t hat s o me p atie nts ma y e x perie nce mil d reacti o ns si milar t o t h ose 
t he y mi g ht e x perie n ce relate d t o Octre o Sca n (a n uclear ( S P E C T) i ma gi n g a ge nt) s u c h as 
dizzi ness, fe ver, c ha n g es i n li ver e nz y mes, a n d na usea. T he d ose escalati o n pr oce d ure will 
all o w us t o m o nit or t hese reacti o ns fr o m l o w t o hi g h d oses of L S 3 0 1. T her e are ot her 
e x peri me ntal t u m or tar gete d i ma gi n g a ge nts c urre ntl y bei n g e val uate d i n h u ma n patie nts  
(see t he ta ble bel o w). M ost of t he m eit her user a nti b o dies ta g ge d wit h I C G  or d ye- la bele d 
bi o m olec ule.  
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 8  of 5 4  
 
 
List of i m a gi n g a ge nts u n der cli nic al i n vesti g ati o n f or i m a ge- g ui de d s ur ger y  
Sl 
n o  
A ge nt  
Cli nic al Tri al  
Tri al t y pe  
C a ncer 
t y pe 
A d verse 
re acti o ns  
1  
Cet u xi m u b 
I R D y e 8 0 0 
N C T 0 1 9 8 7 3 7 5  
P hase 1 d ose 
escalati o n st u d y  
Hea d a n d 
nec k  
Yes, i n 
h u ma ns 6 9 
a n d 
m o n ke ys 7 0 
2  
Pa nit u m u ma b 
I R D y e 8 0 0 
N C T 0 2 4 1 5 8 8 1  
P hase 1 safet y/ 
efficac y st u d y  
Hea d a n d 
nec k  
N o re p orts 
f o u n d 
3  
Be vaciz u ma b -
8 0 0 C W 
N C T 0 1 5 0 8 5 7 2  
N C T 0 2 5 8 3 5 6 8 
P hase 1  
P hase 2  
Breast  
N o re p orts 
f o u n d. 
N o ne 
re p orte d 
f or P E T 
versi o n 7 1 
4  
I C G a n d g o g gle  
N C T 0 2 3 1 6 7 9 5  
Pil ot  
Breast a n d 
Mela n o ma  
N o ne 
re p orte d4 3 
5  
I C G a n d 
Fl u o bea m  
N C T 0 1 7 3 8 2 1 7  
P hase 1 a n d 
P hase 2  
Li ver  
N o re p orts 
f o u n d 
6  
B L Z 1 0 0 a n d 
Fl u o bea m 
N C T 0 2 0 9 7 8 7 5  
P hase 1 d ose 
escalati o n  
S ki n 
ca ncer  
N o ne i n 
h u ma ns 7 2 
a n d d o gs 7 3 
7  
B L Z 1 0 0 a n d 
u ns pecifie d 
s yste m 
N C T 0 2 2 3 4 2 9 7  
P hase 1 safet y 
Gli o ma  
N o ne i n 
h u ma ns 7 2 
a n d d o gs 7 3 
 
T he ris ks fr o m t h e C V G  s yste m are e xtre mel y l o w. T he s yste m d oes n ot c o me i nt o c o ntact 
wit h t he test s u bject. It utilizes safe i n visi ble NI R li g ht bet wee n 7 0 0 a n d 9 0 0 n m at a ver y 
l o w fl ue nce rate ( ≤ 2 5 m W/c m2). T his i ma gi n g s yste m meets t he U S F D A defi niti o n of a 
“ n o n-si g nifica nt ris k” de vice, a n d i n dee d c o ul d be c o nsi dere d “e q ui vale nt ” t o t he S p y ™ 
s yste m, w hic h has alrea d y o btai ne d U S F D A a p pr o val as a n o n-si g nifica nt ris k de vice.  
 
T he U S F D A li mit f or NI R e x p os ure is 1 0, 0 0 0 m W/c m 2 a n d t he c urre nt s yste m utilizes 
2. 5- 2 5 m W/c m 2 f or a t otal d urati o n of less t h a n 3 0 mi n utes i n t he i nitial i ma gi n g a n d 1 0 
mi n utes per s u bse q ue nt i ma gi n g se g me nts. T his is e q ui vale nt t o t he c urre nt NI R d ose 
e mitte d fr o m t he sta n dar d 5 -la nter n s ur gical li g hts c o m m o nl y use d i n t he o perati n g r o o m. 
 
Patie nt safet y m o nit ori n g f or a d v erse e ve nts as a res ult of t his st u d y will be c o n d uct e d as 
descri be d  i n  S ecti o n 5. 4  a b o ve.  A n y  re p orta bl e a d verse  e v e nt  will  be  re p orte d  i n  
acc or d a nce t o Was hi n gt o n U ni versit y H R P O re g ulati o ns  (see Secti o n 6. 0) f or p hase 1 A . 
A n y re p orta ble a d verse e ve nt i n p hase 1 B/ p hase 2 will be re p orte d i n acc or d a nce t o U T S W 
H R P P/I R B g ui deli nes (see Secti o n 6. 0). 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 2 9  of 5 4  
 
 
5. 6  
C ollecti o n of S peci m e ns f or Res e arc h A n al yses  ( P h ase 1 B a n d P h ase 2 
O N L Y)  
 
5. 6. 1  I de ntific ati o n of Bl o o d Pr otei ns t h at Bi n d L S 3 0 1 
 
Fi ve m L  of bl o o d c ollect e d at t he sa me ti me as t h e  C B C a n d C M P at baseli ne, 6 0 
mi n utes p ost -i njecti o n, a n d o ne h o ur bef ore s ur g er y will be use d t o i de ntif y t he 
pre d o mi na nt bl o o d pr otei ns t hat bi n d L S 3 0 1 a n d me diate t he bi oa vaila bilit y of t he 
a ge nt.  T his r esearc h bl o o d will be dra w n i nt o a pi n k t o p li g ht-pr otecte d t u be a n d 
br o u g ht t o t he Ac hilef u l a b. 
 
5. 6. 2  Deter mi n ati o n of L S 3 0 1- Ass oci ate d Bi o m ar kers i n Bre ast C a ncer 
Tiss ue  
 
After  s ur gical  re m o val  of t he ca ncer,  t he sec o n d  ( n o n -o perati n g) s ur ge o n will  
e xa mi ne t he tiss ue wit h t he g o g gles a n d a n a d diti o nal i n k will be use d t o i de ntif y 
a n y L S 3 0 1 fl u oresce nce i n t he s ur gical mar gi n.  T his s ur ge o n will t he n use t he 
g o g gles  t o  pr oc ure  c ores  fr o m  t he  i n ke d  a n d  secti o ne d  s peci me ns  i n  s ur gical  
pat h ol o g y.  T hese c or es will t he n be fr o ze n a n d d eli vere d t o t he bi os peci m e n c ore 
f or accessi o ni n g, pr ocessi n g, a n d st ora ge.  T he s peci me ns will t he n be a nal yze d t o 
i de ntif y L S 3 0 1-ass ociate d bi o mar kers.  
 
5. 7  
D at a S u b missi o n Sc he d ule  
 
Case re p ort f or ms wit h a p pr o priate s o urce d oc u m e ntati o n will be c o m plete d acc or di n g t o t he 
sc he d ule liste d i n t his st u d y. 
 
C ase Re p ort F or m  
S u b missi o n Sc he d ule  
Ori gi nal C o nse nt F or m  
Pri or t o re gistrati o n  
O n St u d y F or m  Me dical Hist or y F or m  
At t he ti me of st u d y e ntr y  
L S 3 0 1 I njecti o n F or m  
After i njecti o n  
S ur ger y F or m  
After s ur g er y  
F oll o w -U p La bs F or m  
3 0 da ys after i njecti o n  
T o xicit y  F or m  
C o nti n u o us  
Deat h F or m  
Ti me of deat h  
Me d Watc h F or m  
As descri be d i n Secti o n 6. 0  
 
5. 7. 1  A d verse E ve nt C ollecti o n i n t he C ase Re p ort F or ms  
 
All a d verse e ve nts t hat o cc ur be gi n ni n g wit h start of treat me nt ( mi n us e x ce pti o ns 
defi ne d i n Secti o n 6. 0) m ust be ca pt ure d i n t he T o xicit y F or m.  Baseli ne A Es 
s h o ul d be ca pt ure d o n t h e Me dical Hist or y F or m. 
 
Partici pa nt deat h d ue t o disease pr o gressi o n s h o ul d be re p ort e d o n t he T o xicit y 
F or m as gr a de 5 disease pr o gressi o n.  If d eat h is d ue t o a n A E (e. g. car diac 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 0  of 5 4  
 
 
dis or ders: car diac arrest), re p ort as a gra d e 5 e ve nt u n der t hat A E.  Partici pa nt 
deat h m ust als o be rec or d e d o n t he Deat h F or m. 
 
5. 8  
W o m e n of C hil d be ari n g P ote nti al 
 
W o me n of c hil d beari n g p ote ntial ( defi ne d as w o me n wit h re g ular me nses, w o me n wit h 
a me n orr h ea, w o me n  wit h  irre g ular  c y cles,  w o m e n  usi n g  a  c o ntrace pti ve  met h o d  t hat  
precl u des wit h dr a wal blee di n g, a n d w o me n w h o h a ve ha d a t u b al li gati o n) are re q uire d t o 
ha ve a n e gati ve ser u m or uri ne pre g n a nc y test wit hi n 7 da ys  pri or t o a d mi nistrati o n of 
L S 3 0 1 bef ore  i ncl usi o n i n t his st u d y.   
 
5. 9  
D ur ati o n of St u d y P artici p ati o n 
 
If at a n y ti me t he c o nstrai nts of t his pr ot oc ol are c o nsi dere d t o be detri me ntal t o t he 
patie nt’s healt h a n d/ or t he patie nt n o l o n ger wis hes t o c o nt i nue i n t he pr ot oc ol, t he patie nt 
s h o ul d be re m o v e d fr o m t he st u d y a n d t h e reas o n f or disc o nti n uati o n d oc u me nte d i n t he 
case r e p ort f or ms . 
 
Patie nts will be o n st u d y f or t he d urati o n of t he scree ni n g pr ocess a n d t he s ur ger y or u ntil 
o ne of t he f oll o wi n g criteria a p plies: 
•  Deat h  
•  A d verse e ve nt(s) t h at, i n t he j u d g me nt of t he i n vesti gat or, ma y ca use s e vere or 
per ma ne nt har m 
•  S us pecte d pre g n a nc y  
•  Seri o us n o nc o m plia nce wit h t he st u d y pr ot oc ol 
•  L ost t o f oll o w- u p 
•  Patie nt wit h dra ws c o nse nt  
•  I n v esti gat or re m o v es t he patie nt fr o m st u d y 
•  T he Site ma n Ca ncer C e nter deci des t o cl ose t he st u d y  f or p hase 1 A  
•  T he U T S W De p art me nt of S ur ger y or Si m m o ns C o m pre he nsi ve Ca ncer Ce nter t o 
cl ose t he st u d y f or p has e 1 B  a n d phase 2  
 
F oll o w- u p will c o nti n ue f or 3 0 da ys p ost-s ur g er y (f or c ollecti o n of f oll o w- u p la b data). 
 
6. 0  
R E G U L A T O R Y A N D R E P O R TI N G R E Q UI R E M E N T S  
 
T he e ntities pr o vi di n g o versi g ht of safet y a n d c o m plia nce wit h t he pr ot oc ol re q uire re p orti n g as 
o utli ne d bel o w.   Please r efer t o A p p e n di x A f or defi niti o ns a n d A p pe n di x B f or a gri d of re p orti n g 
ti meli nes. 
 
A d verse e ve nts will be tr ac ke d f or o ne h o ur f oll o wi n g a d mi nistrati o n of L S 3 0 1.   
•  Baseli ne a d vers e e v e nts, w hic h s hall be rec or de d o n t he me dical hist or y C R F. 
•  F or p ur p os es of t his pr ot oc ol, a d vers e e v e nts c oll ecte d a n d d oc u me nte d o n C R Fs will n ot 
i ncl u de a n y e v e nts relate d t o s ur ger y. 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 1  of 5 4  
 
 
Refer t o t he d ata s u b missi o n sc he d ule i n Secti o n 5. 7  f or i nstr ucti o n o n t he c ollecti o n of A Es i n t he 
E D C.  
 
Re p orti n g re q uire m e nts f or Was hi n gt o n U ni versit y st u d y tea m ma y be f o u n d i n Secti o n 6. 1 a f or 
p hase 1 A .   
 
Re p orti n g re q uire me nts f or U T  S o ut h wester n M e dical Ce nter  st u d y tea m ma y be f o u n d i n Secti o n 
6. 1 b f or p hase 1 B a n d  ph ase 2.   
 
T he Was hi n gt o n U ni versit y H u ma n Researc h Pr otecti o n Office ( H R P O) re q uires t hat all e ve nts 
meeti n g t he defi niti o n of u na ntici pate d pr o ble m or seri o us n o nc o m plia nce be re p orte d as o utli ne d 
i n Secti o n 6. 2 f or p hase 1 A .  
 
T he U T S W H u ma n Researc h Pr otecti o n Pr o gra m Office ( H R P P O) re q uires t hat all e ve nts meeti n g 
t he defi niti o n of u na ntici pate d pr o ble m  or s eri o us n o nc o m plia nce be re p orte d as o utli ne d i n 
Secti o n 6. 2 f or p hase 1 B a n d  phase 2.  
 
T he F D A re q uires t hat all seri o us a n d u ne x pect e d a d verse e v e nts be re p ort e d as o utli ne d i n Secti o n 
6. 4.  I n a d diti o n, a n y fatal or life -t hreate ni n g a d verse e x p erie n ces w h ere t here is a reas o na bl e 
p ossi bilit y of relati o ns hi p t o st u d y i nter ve nti o n m ust be re p orte d. 
 
F or t he p ur p oses of t his st u d y, A Es relat e d t o s ur ger y will n ot be c ollect e d or c o nsi dere d re p ort a ble.  
 
6. 1  
S p o ns or -I n vesti g at or Re p orti n g Re q uire me nts – W as hi n gt o n U ni versit y 
 
P h ase 1 A: W as hi n gt o n U ni versit y  
 
6. 1. 1  Re p orti n g t o t he H u m a n Rese arc h Pr otecti o n Office ( H R P O) at 
W as hi n gt o n U ni versit y 
 
Re p orti n g will be c o n d ucte d i n acc or d a nce wit h Was hi n gt o n U ni versit y I R B 
P olicies.  
 
Pre -a p pr o v al of all pr ot o c ol e xce pti o ns m ust be o btai ne d pri or t o i m ple me nti n g 
t he c ha n ge. 
 
6. 1. 2  Re p orti n g t o t he Q u alit y Ass ur a nce a n d S afet y M o nit ori n g 
C o m mittee ( Q A S M C) at W as hi n gt o n U ni versit y  
 
T he  S p o ns or  I n vesti gat or  ( or  desi g nee)  is  re q uire d  t o  n otif y  Q A S M C  of  a n y  
u na ntici pate d pr o ble ms i n v ol vi n g ris ks t o partici pa nts or ot hers occ urri n g at W U or 
a n BJ H or S L C H i nstit uti o n t hat ha ve bee n re p orte d t o a n d ac k n o wle d ge d b y 
H R P O.  ( U na ntici pate d pr o ble ms re p orte d t o H R P O a n d wit h dra w n d uri n g t he 
re vie w pr o cess nee d n ot be re p ort e d t o Q A S M C.) 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 2  of 5 4  
 
 
Q A S M C m ust be n otifie d wit hi n 1 0 d a ys  of recei pt of I R B ac k n o wle d ge m e nt via 
e mail t o qas mc @ w ustl.e d u .  S u b missi o n t o Q A S M C m ust i ncl u de t he m yI R B f or m 
a n d a n y s u p p orti n g d oc u me ntati o n se nt wit h t he f or m. 
 
6. 1. 3  Re p orti n g t o t he F D A  
 
T he c o n d uct of t he st u d y will c o m pl y wit h all F D A safet y re p orti n g re q uire me nts.  
P L E A S E N O T E T H A T R E P O R TI N G R E Q UI R E M E N T S F O R T H E F D A 
DI F F E R F R O M R E P O R TI N G R E Q UI R E M E N T S F O R H R P O/ Q A S M C.  It 
is t he res p o nsi bilit y of t he i n vesti gat or t o re p ort a n y u na ntici pate d pr o ble m t o t he 
F D A as f oll o ws:  
•  Re p ort a n y u n e x pect e d fatal or life -t hreate ni n g s us pecte d a d vers e reacti o n 
(ref er t o A p pe n di x A f or defi niti o ns) n o later t ha n 7 c ale n d ar d a ys  after 
i nitial recei pt of t he i nf or mati o n.   
•  Re p ort a s us pecte d a d verse reacti o n t hat is b ot h seri o us a n d u ne x pecte d 
( S U S A R, refer t o A p pe n di x A), n o later t ha n 1 5 c ale n d ar d a ys  after it is 
deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g.  Re p ort a n a d v erse 
e ve nt (refer t o A p pe n di x A) as a s us pecte d a d v ers e reacti o n o nl y if t here is 
e vi de nce t o s u g gest a ca usal relati o ns hi p bet w ee n t he dr u g a n d t he a d v erse 
e ve nt, s uc h as: 
o  A si n gle occ urre nce of a n e ve nt t hat is u nc o m m o n a n d k n o w n t o be 
str o n gl y ass ociate d wit h dr u g e x p os ure 
o  O ne or m ore o cc urre n ces of a n e ve nt t hat is n ot c o m m o nl y ass ociate d 
wit h dr u g e x p os ure b ut is ot her wise u nc o m m o n i n t he p o p ulati o n 
e x p ose d t o t he dr u g 
o  A n a g gre g ate a n al ysis of s pecific e v e nts o bser ve d i n a cli nical trial t hat 
i n dicates t h ose e ve nts occ ur m ore fre q u e ntl y i n t he dr u g treat me nt 
gr o u p t ha n i n a c o nc urre nt or hist orical c o ntr ol gr o u p 
•  Re p ort a n y fi n di n gs fr o m e pi de mi ol o gical st u dies, p o ole d a nal ysis of 
m ulti ple st u dies, or cli nical st u dies t hat s u g gest a si g nifica nt ris k i n 
h u ma ns e x p ose d t o t he dr u g n o later t ha n 1 5 c ale n d ar d a ys  after it is 
deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g. 
•  Re p ort a n y fi n di n gs fr o m a ni mal or i n vitr o testi n g t hat s u g gest si g nifica nt 
ris k i n h u ma ns e x p ose d t o t he dr u g n o later t ha n 1 5 cale n dar da ys after it is 
deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g.  
•  Re p ort a n y cli nicall y i m p orta nt i ncrease i n t he rate of a seri o us s us pect e d 
a d verse reacti o n of t hat liste d i n t he pr ot oc ol or I B wit hi n 1 5 cale n d ar d a ys 
after it is deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g.  
 
S u b mit eac h r e p ort as a n I N D safet y re p ort i n a n arrati ve f or m at or o n F D A F or m 
3 5 0 0 A or i n a n electr o nic f or mat t hat F D A ca n pr o cess, re vie w, a n d arc hi ve.  St u d y 
tea ms m ust n otif y t he Site ma n Ca ncer Ce nter Pr ot oc ol De vel o p me nt tea m of eac h 
p ote ntiall y  re p orta ble  e ve nt  wit hi n  1  b usi ness  da y  after  i nitial  recei pt  of  t he  
i nf or mati o n, a n d m ust bri n g t he si g ne d 1 5 7 1 a n d F D A F or m 3 5 0 0 A t o t he Site ma n 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 3  of 5 4  
 
 
Ca ncer Ce nt er Pr ot oc ol De vel o p me nt tea m n o lat er t ha n 1 b usi ness d a y pri or t o t he 
d ue date f or re p orti n g t o t he F D A. 
 
Eac h n otificati o n t o F D A m ust bear pr o mi ne nt i de ntificati o n of its c o nte nts (“I N D 
Safet y Re p ort”) a n d m ust be tra ns mitte d t o t h e re vie w di visi o n i n t he Ce nter f or 
Dr u g E val u ati o n a n d Researc h ( C D E R) or i n t h e Ce nter f or Bi ol o gics E v al uati o n 
a n d Researc h ( C B E R) t hat has res p o nsi bilit y f or re vie w of t he I N D.  R ele va nt 
f oll o w-u p i nf or mati o n t o a n I N D safet y re p ort m ust be s u b mitte d as s o o n as t he 
i nf or mati o n is a vaila bl e a n d m ust be i de ntifie d as s uc h (“ F oll o w-u p I N D Safet y 
Re p ort”).  
 
6. 2  
S p o ns or -I n vesti g at or Re p orti n g Re q uire me nts – U T S o ut h wester n  
 
P h ase 1 B a n d P h ase 2:  
 
6. 2. 1  Re p orti n g t o t he H u m a n Rese arc h Pr otecti o n Pr o gr a m office 
( H R P P) at U T S o ut h wester n Me dic al C e nter 
 
Re p orti n g will be c o n d ucte d i n acc or d a nce wit h U T S o ut h wester n Me dical Ce nter 
I R B P olicies. 
 
Pre -a p pr o v al of all pr ot o c ol e xce pti o ns m ust be o btai ne d pri or t o i m ple me nti n g 
t he c ha n ge. 
 
6. 2. 2  Re p orti n g t o t he D at a a n d S afet y M o nit ori n g C o m mittee ( D S M C) at 
U T S o ut h wester n Me di c al Ce nter  
 
F or  l o cal  p atie nts,  i n  a d diti o n  t o  t he  s p o ns or  m o nit ori n g,  t he  Si m m o ns  
C o m pre he nsi ve Ca n cer Data Safet y a n d M o nit ori n g C o m mittee ( D S M C) will be 
m o nit ori n g o nl y l ocal u n a ntici pate d S A E’s as re p orte d t o t he I R B. T he c h air will 
re vie w  o nl y  l ocal  u na ntici pate d S A E’s  i n  real  ti me.  T hese  are  t he n  re vie we d  
q uarterl y at D S M C meeti n gs T he c hair ma n of t h e D S M C is res p o nsi ble f or t his 
o n g oi n g m o nit ori n g. M o nit ori n g of seri o us a d verse e ve nts ha p pe ns b y t he D S M C 
c hair ma n i n real ti me a n d a n a n n ual re vi e w ta kes place t o l o o k at tre n ds i n a d verse 
e ve nts, accr ual a n d ot her a p plica ble st u d y e ve nts. T he Pri nci pal I n vesti gat or ( or a 
desi g nate d  c o -i n v esti gat or)  will  be  pri maril y  res p o nsi ble  f or  m o nit ori n g  st u d y  
accr u al rate, st u d y attriti o n/ wit h dra wals/ dr o p o uts, patter ns of a d v erse e ve nts a n d/ or 
u na ntici pate d e ve nt a n d patter ns of pr ot oc ol vi olati o ns a n d/ or de viati o ns at t his site.  
 
Trial m o nit ori n g will be c o n d ucte d n o l ess t ha n a n n uall y a n d refers t o a re g ular 
i nter val re vi e w of trial rel ate d acti vit y a n d d oc u me ntati o n w hic h i ncl u des b ut is n ot 
li mite d  t o  acc urac y  of  case  re p ort  f or ms,  pr ot o c ol  c o m plia nce,  ti meli ness  a n d  
acc ur ac y of  Vel os e ntri es a n d A E/ S A E ma na ge me nt a n d re p orti n g. D oc u me ntati o n 
of trial m o nit ori n g will be mai ntai ne d al o n g wit h ot her pr ot oc ol relate d d o c u me nts 
a n d will be re vie w e d d uri n g i nter nal a u dit.   
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 4  of 5 4  
 
 
T he  S p o ns or  I n vesti gat or  ( or  desi g nee)  is  re q uire d  t o  n otif y  D S M C  of  a n y  
u na ntici pate d pr o ble ms i n v ol vi n g ris ks t o partici pa nts or ot hers occ urri n g at U T S W 
t hat ha ve bee n r e p orte d t o a n d ac k n o wle d ge d b y H R P P.  ( U na ntici pate d pr o ble ms 
re p orte d t o H R P P a n d wit h dra w n d uri n g t he re vie w pr ocess nee d n ot be re p orte d t o 
D S M C.)  
 
D S M C m ust be n otifie d i n real -ti me as t he y are re p orte d t o t he I R B t o t he U T S W 
Si m m o ns Ca ncer Ce nter D S M C C o or di nat or.  
 
6. 2. 3  Re p orti n g t o t he F D A  
 
T he c o n d uct of t he st u d y will c o m pl y wit h all F D A safet y re p orti n g re q uire me nts.  
P L E A S E N O T E T H A T R E P O R TI N G R E Q UI R E M E N T S F O R T H E F D A 
DI F F E R F R O M R E P O R TI N G R E Q UI R E M E N T S F O R H R P P/ D S M C.  It is 
t he res p o nsi bilit y of t h e i n vesti gat or t o re p ort a n y u na ntici pate d pr o ble m t o t he 
F D A as f oll o ws:  
•  Re p ort a n y u n e x pect e d fatal or life -t hreate ni n g s us pecte d a d vers e reacti o n 
(ref er t o A p pe n di x A f or defi niti o ns) n o later t ha n 7 c ale n d ar d a ys  after 
i nitial recei pt of t he i nf or mati o n.   
•  Re p ort a s us pecte d a d verse reacti o n t hat is b ot h seri o us a n d u ne x pecte d 
( S U S A R, refer t o A p pe n di x A), n o later t ha n 1 5 c ale n d ar d a ys  after it is 
deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g.  Re p ort a n a d v erse 
e ve nt (refer t o A p pe n di x A) as a s us pecte d a d v ers e reacti o n o nl y if t here is 
e vi de nce t o s u g gest a ca usal relati o ns hi p bet w ee n t he dr u g a n d t he a d v erse 
e ve nt, s uc h as: 
o  A si n gle occ urre nce of a n e ve nt t hat is u nc o m m o n a n d k n o w n t o be 
str o n gl y ass ociate d wit h dr u g e x p os ure 
o  O ne or m ore o cc urre n ces of a n e ve nt t hat is n ot c o m m o nl y ass ociate d 
wit h dr u g e x p os ure b ut is ot her wise u nc o m m o n i n t he p o p ulati o n 
e x p ose d t o t he dr u g 
o  A n a g gre g ate a n al ysis of s pecific e v e nts o bser ve d i n a cli nical trial t hat 
i n dicates t h ose e ve nts occ ur m ore fre q u e ntl y i n t he dr u g treat me nt 
gr o u p t ha n i n a c o nc urre nt or hist orical c o ntr ol gr o u p 
•  Re p ort a n y fi n di n gs fr o m e pi de mi ol o gical st u dies, p o ole d a nal ysis of 
m ulti ple st u dies, or cli nical st u dies t hat s u g gest a si g nifica nt ris k i n 
h u ma ns e x p ose d t o t he dr u g n o later t ha n 1 5 c ale n d ar d a ys  after it is 
deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g. 
•  Re p ort a n y fi n di n gs fr o m a ni mal or i n vitr o testi n g t hat s u g gest si g nifica nt 
ris k i n h u ma ns e x p ose d t o t he dr u g n o later t ha n 1 5 cale n dar da ys after it is 
deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g.  
•  Re p ort a n y cli nicall y i m p orta nt i ncrease i n t he rate of a seri o us s us pect e d 
a d verse reacti o n of t hat liste d i n t he pr ot oc ol or I B wit hi n 1 5 cale n d ar d a ys 
after it is deter mi ne d t hat t he i nf or mati o n q ualifies f or re p orti n g.  
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 5  of 5 4  
 
 
S u b mit eac h r e p ort as a n I N D safet y re p ort i n a n arrati ve f or m at or o n F D A F or m 
3 5 0 0 A or i n a n electr o nic f or mat t hat F D A ca n pr o cess, re vie w, a n d arc hi ve.  St u d y 
tea ms m ust n otif y t he D e part me nt of S ur ger y Researc h tea m or Si m m o ns Ca ncer 
Ce nter tea m of ea c h p ot e ntiall y re p orta ble e v e nt wit hi n 1 b usi ness da y after i nitial 
recei pt of t he i nf or mati o n, a n d m ust bri n g t he si g ne d 1 5 7 1 a n d F D A F or m 3 5 0 0 A 
t o t he De part me nt of S ur ger y Researc h tea m or Si m m o ns Ca ncer Ce nter tea m n o 
later t ha n 1 b usi ness da y pri or t o t he d ue date f or re p orti n g t o t he F D A. 
 
Eac h n otificati o n t o F D A m ust bear pr o mi ne nt i de ntificati o n of its c o nte nts (“I N D 
Safet y Re p ort”) a n d m ust be tra ns mitte d t o t h e re vie w di visi o n i n t he Ce nter f or 
Dr u g E val u ati o n a n d Researc h ( C D E R) or i n t h e Ce nter f or Bi ol o gics E v al uati o n 
a n d Researc h ( C B E R) t hat has res p o nsi bilit y f or re vie w of t he I N D.  R ele va nt 
f oll o w-u p i nf or mati o n t o a n I N D safet y re p ort m ust be s u b mitte d as s o o n as t he 
i nf or mati o n is a vaila bl e a n d m ust be i de ntifie d as s uc h (“ F oll o w-u p I N D Safet y 
Re p ort”).  
 
6. 3  
E xce pti o ns t o E x pe dite d Re p orti n g  – W as hi n gt o n U ni versit y a n d U T 
S o ut h wester n  
 
E ve nts t hat d o n ot re q uir e e x pe dite d re p orti n g as descri be d i n Secti o n 6. 1 i ncl u de: 
•  pla n ne d h os pitalizati o ns 
•  h os pitalizati o ns < 2 4 h o urs 
•  res pite care 
•  e ve nts relat e d t o disease pr o gressi o n  
 
E ve nts t hat d o n ot re q uir e e x pe dite d re p orti n g m ust still be ca pt ure d i n t he E D C.  
 
 
7. 0  
D A T A A N D S A F E T Y M O NI T O RI N G  
 
7. 1  
P h ase 1 A: W as hi n gt o n U ni versit y 
I n c o m plia nce wit h t he Was hi n gt o n U ni versit y I nstit uti o nal Data a n d Safet y M o nit ori n g Pla n, t he 
Pri nci pal I n vesti gat or will pr o vi de a Data a n d Safet y M o nit ori n g ( D S M) re p ort t o t he Was hi n gt o n 
U ni versit y  Q ualit y  Ass ura nce  a n d  Safet y  M o nit ori n g  C o m mittee  ( Q A S M C)  se mi -a n n uall y 
be gi n ni n g si x m o nt hs aft er accr ual has o pe n e d (if at least o ne p atie nt has bee n e nr olle d) or o n e 
year after accr ual has o p e ne d (if n o patie nts ha v e b ee n e nr olle d at t he si x - m o nt h mar k).   
 
T he Pri nci pal I n vesti gat or will re vie w all patie nt data at least e ver y 6 m o nt hs, a n d pr o vi de a se mi -
a n n ual re p ort t o t he Q A S M C.  T his re p ort will i ncl u de:  
•  H R P O pr ot oc ol n u m ber, pr ot oc ol title, Pri nci pal I n vesti gat or na me, data c o or di nat or na me, 
re g ulat or y c o or di nat or na me, a n d statisticia n na me  
•  Date of i nitial H R P O a p pr o val, date of m ost rece nt c o nse nt H R P O a p pr o v al/re visi o n, date 
of H R P O e x pirati o n, date of m ost rece nt Q A a u dit, st u d y stat us, a n d p hase of st u d y 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 6  of 5 4  
 
 
•  Hist or y of st u d y i ncl u di n g s u m mar y of s u bsta nti ve a me n d me nts; s u m mar y of accr u al 
s us pe nsi o ns i ncl u di n g start/st o p dates a n d reas o n; a n d s u m mar y of pr ot o c ol e xce pti o ns, 
err or, or breac h of c o nfi d e ntialit y i ncl u di n g start/st o p dates a n d reas o n 
•  St u d y- wi de tar get accr u al a n d st u d y -wi de act u al accr ual  
•  Pr ot oc ol acti vati o n date  
•  A vera g e rat e of accr u al o bser ve d i n year 1, year 2, a n d s u bse q ue nt years 
•  E x pecte d accr ual e n d dat e a n d accr u al b y c o h ort 
•  O bjecti ves of pr ot oc ol wit h s u p p orti n g data a n d list t he n u m ber of p artici p a nts w h o ha ve 
met eac h o bjecti ve  
•  Earl y st o p pi n g r ules wit h s u p p orti n g data a n d list t he n u m ber of partici pa nts w h o ha ve met 
t he earl y st o p pi n g r ules  
•  S u m mar y of t o xicities s e parat e d b y c o h orts wit h t he n u m ber of d ose-li miti n g t o xicities 
i n dicate d 
•  A bstract s u b missi o ns/ p u blicati o ns 
•  S u m mar y of a n y rece nt literat ure t h at ma y affect t he safet y or et hics of t he st u d y  
 
T he  st u d y  pri nci pal  i n vesti gat or  a n d  Researc h  Patie nt  C o or di nat or  will  m o nit or  f or  seri o us  
t o xicities o n a n o n g oi n g basis. O nce t he pri nci pal i n vesti gat or or Researc h Patie nt C o or di nat or 
bec o mes a war e of a n a d v erse e ve nt, t he A E will b e re p orte d t o t he H R P O a n d Q A S M C acc or di n g 
t o i nstit uti o nal g ui deli nes. 
 
7. 2  
P h ase 1 B a n d P h ase 2: U T S W  
I n c o m plia nce wit h t he Si m m o ns Ca ncer Ce nter Data a n d Safet y M o nit ori n g C o m mittee, t he 
Pri nci pal I n vesti gat or will pr o vi de a Data a n d Safet y M o nit ori n g ( D S M) re p ort t o t he Si m m o ns 
Ca ncer Ce nter Data a n d Safet y M o nit ori n g C o m mittee ( D S M C) se mi -a n n uall y be gi n ni n g si x 
m o nt hs after accr u al has o pe ne d (if at least o ne patie nt has bee n e nr olle d) or o ne year after accr ual 
has o pe ne d (if n o patie nts ha ve bee n e nr olle d at t he si x- m o nt h mar k).    
 
T he Pri nci pal I n vesti gat or will re vie w all patie nt data at least e ver y 6 m o nt hs, a n d pr o vi de a se mi -
a n n ual re p ort t o t he D S M C.  T his re p ort will i ncl u de: 
•  H R P P pr ot oc ol n u m ber, pr ot oc ol title, Pri nci pal I n vesti gat or na me, data c o or di nat or na me, 
re g ulat or y c o or di nat or na me, a n d statisticia n na me 
•  Date of i nitial H R P P a p pr o val, date of m ost rece nt c o nse nt H R P P a p pr o val/re visi o n, date 
of H R P P e x pirati o n, date of m ost rece nt Q A a u dit, st u d y stat us, a n d p hase of st u d y 
•  Hist or y of st u d y i ncl u di n g s u m mar y of s u bsta nti ve a me n d me nts; s u m mar y of accr u al 
s us pe nsi o ns i ncl u di n g start/st o p dates a n d reas o n; a n d s u m mar y of pr ot o c ol e xce pti o ns, 
err or, or breac h of c o nfi d e ntialit y i ncl u di n g start/st o p dates a n d reas o n 
•  St u d y- wi de tar get accr u al a n d st u d y -wi de act u al accr ual  
•  Pr ot oc ol acti vati o n date  
•  A vera g e rat e of accr u al o bser ve d i n year 1, year 2, a n d s u bse q ue nt years 
•  E x pecte d accr ual e n d dat e a n d accr u al b y c o h ort 
•  O bjecti ves of pr ot oc ol wit h s u p p orti n g data a n d list t he n u m ber of p artici p a nts w h o ha ve 
met eac h o bjecti ve  
•  Earl y st o p pi n g r ules wit h s u p p orti n g data a n d list t he n u m ber of partici pa nts w h o ha ve met 
t he earl y st o p pi n g r ules  

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 7  of 5 4  
 
 
•  S u m mar y of t o xicities s e parat e d b y c o h orts wit h t he n u m ber of d ose -li miti n g t o xicities 
i n dicate d 
•  A bstract s u b missi o ns/ p u blicati o ns 
•  S u m mar y of a n y rece nt literat ure t h at ma y affect t he safet y or et hics of t he st u d y  
 
T he st u d y pri nci pal i n vesti gat or a n d Researc h St u d y C o or di nat or will m o nit or f or seri o us t o xicities 
o n a n o n g oi n g basis. O nce t he pri nci pal i n vesti gat or or Researc h St u d y C o or di nat or bec o mes 
a war e  of  a n  a d verse  e v e nt,  t he  A E  will  be  re p orte d  t o  t he  H R P P  a n d  D S M C  acc or di n g  t o  
i nstit uti o nal g ui deli nes.  
 
8. 0  
S T A TI S TI C A L C O N SI D E R A TI O N S  
 
8. 1  
St u d y D esi g n 
 
A r olli n g si x desi g n w hi c h is i nte n de d f or relati v el y safe trial s a n d s h orte ni n g t he st u d y 
d urati o n is pr o p ose d f or t he p hase 1 p orti o n 7 4. I n t he r olli n g si x desi g n, three  t o si x patie nts 
ca n be e nr olle d t o a d ose eac h ti me. T h e d ose d e -escalati o n occ urs w he n t w o or m ore D L T 
occ ur at a d ose le v el w hile d ose escalati o n occ urs w he n 3/ 3, 4/ 4, 5/ 5, 5/ 6 or 6/ 6 patie nts 
are e val uate d wit h o ut D L T  7 5. T hree d ose le v els will be teste d i n t he p h ase 1 st u d y, 0. 05, 
0. 0 7 5 a n d 0. 10 m g/ k g. L S 3 0 1 will be a d mi nistere d i ntra ve n o usl y at a starti n g d ose of 0. 0 5 
m g/ k g t o a c o h ort of si x patie nts a n d escalate/ de -es calate f oll o wi n g t he decisi o n r ule of t he 
r olli n g si x desi g n. O nce si x patie nts ha ve bee n i ncl u de d at t he c urre nt d ose l e vel, i ncl usi o ns 
are s us pe n de d u ntil at least fi ve of t he si x patie nts  ha ve c o m plete d t he pr oce d ure wit h o ut 
D L T o bser ve d i n t he e val uati o n peri o d. M T D is hit w here t w o or m ore patie nts o ut of si x 
at a d ose le vel e x perie nce D L T . We d o n ot e x pect a n y seri o us a d verse e ve nts relate d t o 
L S 3 0 1 at t h e i nitial D ose Le vel 1. O nce  t he M T D is deter mi ne d, a n e x pa nsi o n c o h ort wit h 
9 patie nts will be teste d at t he M T D a n d an o pti mal i ma gi n g d ose will be rec o m me n de d f or 
t he s u bse q ue nt p hase 2 trial.  
 
At t he c o m pleti o n of t he e x pa nsi o n c o h ort , a si n gl e ar m p h ase 2 trial will b e perf or me d at 
t he o pti mal i ma gi n g d os e. A breast s ur g e o n will perf or m breast c o nser vi n g s ur ger y per 
sta n dar d of care. At t he c o m pleti o n of t he s ur ger y, a sec o n d i n vesti gat or (t o re d uce bias i n 
data c ollecti o n) will wear t he ca ncer visi o n g o g gle ( C V G) t o vis ualize t he e xcise d tiss ue t o 
deter mi ne if t h ere ar e a n y p ositi ve mar gi ns base d o n t he prese nce of L S 3 0 1 fl u oresce nce. 
If a n y p ositi ve mar gi ns are o bser v e d, t he s u bject will be c o nsi dere d t o ha ve a p ositi ve 
mar gi n; ot her wise, t h e mar gi n is c o nsi dere d ne gati ve. T he e xcise d tiss ue will later be 
e xa mi ne d b y  a breast ca ncer pat h ol o gist. Sta n dar d pat h ol o gic tec h ni q ues will be use d t o 
deter mi ne if t here are p ositi ve mar gi ns. Partic ular atte nti o n will be pai d t o t he sites mar ke d 
f oll o wi n g C V G ass ess me nt t o c o nfir m w het her t he mar gi n is p ositi ve or n e g ati ve at a ny of 
t hese sites, or p ositi ve at ot her l ocati o ns n ot i de ntifie d b y C V G. N o i nter v e nti o ns will be 
all o we d base d o n t he C V G res ults. 
    
8. 2  
E n d p oi nts  
 
8. 2. 1  E n d p oi nts f or S afet y A n al ysis 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 8  of 5 4  
 
 
T he pri mar y e n d p oi nts of t he p hase 1 p orti o n of t he trial are t o xicities a n d/ or 
a d verse e v e nts (as o utli ne d i n t he Nati o nal Ca ncer I nstit ute C o m m o n Ter mi n ol o g y 
Crite ria f or A d vers e E ve nts versi o n 5. 0) rec or de d after L S 3 0 1 i njecti o n w hic h are 
defi niti vel y or p ossi bl y relate d t o t he i njecti o n.  Safet y e val uati o n i ncl u des vital 
si g ns, cli nical la b orat or y testi n g a n d E C G, meas ure d pre- ( wit hi n 1 0-1 5 mi n utes or 
3 0  mi n utes  of  i njecti o n)  a n d  p ost -i njecti o n  ( wit hi n  3 0  mi n utes  or/a n d  at  ~ 6 0  
mi n utes). N ote w ort h y p ost - vers us-pre c ha n g es i n all t he safet y meas ures will be 
rec or d ed w hile cli nicall y si g nifica nt c ha n ges will be re p orte d as a d verse e v e nts i n 
acc or d a nce t o H R P O  re q uire me nts. All t he safet y e n d p oi nts are d escri be d i n detail 
i n Secti o n 5. 4 a n d a d vers e e ve nts detaile d  i n Secti o n 6. 0.  
 
8. 2. 2  E n d p oi nt f or P re dicti o n of P ositi ve M ar gi n a n d P ositi ve S L N 
A n al ysis 
 
T he pri mar y o bjecti ve is t o assess mar gi n stat us calle d b y L S 3 0 1 a n d C V G i n 
c o m paris o n t o t he g ol d sta n dar d hist o pat h ol o gical res ults. T he hist o pat h ol o gical 
res ults of mar gi n p ositi vit y has t he f oll o wi n g p ossi ble res ults: mar gi n ne gati ve, 
mar gi n p ositi ve at L S 3 0 1 a n d C V G i de ntifie d l ocati o ns, mar gi n p ositi ve at L S 3 0 1 
a n d C V G u ni de ntifie d l ocati o ns. Usi n g L S 3 0 1 a n d C V G, t he e xercise d tiss ue will 
be c o nsi der e d t o be m ar gi n p ositi ve a n d ha ve i n k mar ke d at t h e i de ntifie d l ocati o ns 
if a n y p ositi ve mar gi ns are o bser ve d; ot her wise, t he mar gi n is c o nsi dere d ne gati ve. 
Dia g n ostic test c haracteristics (se nsiti vit y a n d s pecificit y) of L S 3 0 1 a n d C V G f or 
mar gi n p ositi vit y b y hist ol o g y will esta blis he d a n d mar gi n p ositi vit y rate, defi ne d 
as t he pr o p orti o n of t u m ors wit h hist o pat h ol o gicall y c o nfir me d p ositi ve mar gi ns 
b ut misse d b y C V G, will be esti mate d. T he sec o n d ar y e n d p oi nt of t he p hase 2 trial 
is t o xicit y.  
 
8. 3  
D at a A n al ysis 
 
Cli nical a n d de m o gra p hic c haracteristics of t he partici pa nts will be s u m marize d usi n g 
descri pti ve statistics, o verall a n d b y d ose le v els. 
 
8. 3. 1  S afet y A n al ysis   
 
A d verse e ve nts will be s u m marize d b y descri pti ve statistics (c o u nts, perce nta ges) 
o verall a n d b y d ose le v el  (if a p plica ble).  T h e p ost-i njecti o n vital si g ns will be 
se parat el y c o m pare d t o t he pre-i njecti o n meas ure me nts b y t w o-sa m ple t-test or 
Wilc o x o n ra n k s u m test as a p pr o pri ate, acr oss all patie nts a n d b y d ose le vel  (if 
a p plica ble).  
 
8. 3. 2  M ar gi n P ositi vit y a n d S L N P ositi vit y P re dicti o n A n al ysis 
 
Mar gi n p ositi vit y as deter mi ne d b y L S 3 0 1 a n d C V G will be t a b ulate d wit h t he g ol d 
sta n dar d hist o pat h ol o gical e xa mi nati o n res ults as t he ta ble s h o w n bel o w. 
  
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 3 9  of 5 4  
 
 
  
  
Hist o p at h ol o g y res ults  
  
  
Ne gati ve  
P ositi ve at 
C V G i de ntifie d 
l ocati o ns 
P ositi ve at ot her 
u ni de ntifie d 
l ocati o ns 
C V G 
res ults  
Ne gati ve  
n 1 1 
0  
n 1 3 
P ositi ve at C V G 
i de ntifie d l ocati o ns 
n 2 1 
n 2 2 
0  
 
Dia g n ostic test o perati n g c haracteristics i ncl u di n g se nsiti vit y, s pecificit y, o verall 
acc ur ac y,  p ositi ve  a n d  ne gati ve  pre dicti ve  val u e  will  be  calc ulate d.  T he  tr ue  
p ositi ves ( = n 2 2) are t he t u m ors w hic h s h o w p ositi ve mar gi ns at a n y l ocati o n a gree d 
b y b ot h t he hist o pat h ol o g y e xa mi nati o n a n d b y L S 3 0 1 a n d C V G. S pecificit y is 
defi ne d as t he pr o p orti o n of t u m ors wit h hist o pat h ol o gicall y c o nfir me d n e gati ve 
mar gi ns w hic h are calle d t o ha ve  ne g ati ve mar gi ns b y L S 3 0 1 a n d C V G, calc ulate d 
as n 1 1/(n 1 1+  n 2 1). Se nsiti vit y is defi ne d as t he pr o p orti o n of t u m ors w hi c h are 
pre dicte d t o ha ve p ositi ve mar gi ns b y L S 3 0 1 a n d C V G a m o n g t he t u m ors wit h 
hist o pat h ol o gica ll y c o nfir me d p ositi ve mar gi ns b ot h at C V G i de ntifie d l ocati o ns 
a n d at ot her l ocati o ns u ni de ntifie d b y C V G, cal c ulate d as n 2 2/(n 1 3+ n 2 2). O verall 
acc ur ac y is defi ne d as t h e pr o p orti o n of t u m ors wit h mar gi n p ositi vit y/ ne gati vit y 
c orrectl y  pre dicte d  b y  t he  C V G  a m o n g  all  t u m ors,  calc ulate d  as  ( n 1 1+  n 2 2)/( 
n 1 1+ n 1 3+ n 2 1+ n 2 2). P ositi ve pre di cti ve val u e is defi ne d as t he pr o p orti o n of t u m ors 
wit h  mar gi ns  calle d  p ositi ve  at  L S 3 0 1  a n d  C V G  i de ntifie d  l ocati o ns  b y  t he  
hist o pat h ol o g y e xa mi nati o n a m o n g all t h ose pre di cte d t o ha ve p ositi ve mar gi ns b y 
C V G, calc ulat e d as n 2 2/(n 2 1+ n 2 2). Ne gati ve pre dicti ve val ue w as defi n e d as t he 
pr o p orti o n of t u m ors wit h tr ul y ne gati ve mar gi ns a m o n g t h ose pre dict e d s o b y 
C V G, calc ulate d as  n1 1/(n 1 1+ n 1 3). T he mar gi n p ositi vit y rate base d o n L S 3 0 1 a n d 
C V G  will  be  esti mate d  as  n 1 3/(  n1 1+ n 1 3+  n 2 1+ n 2 2).  All  t he  esti mates  will  be  
acc o m pa ni e d  wit h  9 5 %  c o nfi de n ce  i nter val.  Mar gi n  p ositi vit y  rate  will  be  
c o m pare d a g ai nst t he c urre nt 1 5. 5 % mar gi n p ositi vit y rate usi n g t he t w o -si de d o ne-
sa m ple bi n o mial e xact test. Gi ve n e n o u g h D CI S a n d i n vasi ve breast ca n cer (I B C), 
si milar dia g n ostic c har acteristics will be calc ulat e d se paratel y i n t he s u bset t u m ors 
of D CI S a n d I B C.  
 
8. 4  
S a m ple Size  
 
T he sa m ple size f or t he p hase 1 part  will de pe n d o n t he o bser ve d D L T at eac h d ose le vels . 
T he p hase 1 e x pa nsi o n c o h ort will use 9 e val ua ble patie nts. T he ta ble bel o w pr o vi des t he 
pr o ba bilit y of o bser vi n g D L T i n > = 3 3 % i n 9 patie nts  base d o n t he bi n o mial distri b uti o n . If 
t he tr ue D L T rate is as l o w as 5%, s uc h a pr o ba bilit y is s mall, o nl y 0. 0 0 8 4 b ut if t he tr ue 
D T L is 5 0 %, s uc h a pr o b a bilit y is ver y hi g h at 0. 9 1. 
 
 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 4 0  of 5 4  
 
 
T rue D L T  
pr o ba bilit y of o bser ve d D L T > = 3 3 % i n 9 
patie nts  
1 %  
0. 0 0 0 0 8 4  
5 %  
0. 0 0 8 4  
1 0 %  
0. 0 5 3  
2 0 %  
0. 2 6 2  
3 0 %  
0. 5 3 7  
4 0 %  
0. 7 6 8  
5 0 %  
0. 9 1  
 
T he sa m ple size f or t he p hase 2 p orti o n of t he st u d y was deter mi ne d usi n g t he f oll o wi n g 
calc ulati o ns. A t otal of 8 8 ( 1 4 are e x pecte d t o ha ve p ositi ve mar gi ns after sta n dar d  breast 
s ur ger y) si m ulta ne o usl y ac hie ves 8 4. 7 % p o wer t o detect a n e x p ecte d s e nsiti vit y of 0. 9 5 
a gai nst  a n u nacce pta ble s e nsiti vit y of 0. 7 a n d 1 0 0 % p o wer t o detect a n e x p ecte d s pecificit y 
0. 9 5 a gai nst a n u nfa v or a ble s pecificit y of 0. 7 usi n g a o ne -si de d bi n o mial test. T he tar g et 
si g nifica nce le vel is 0. 0 5. T he act ual si g nifica nce le vel ac hie ve d b y t he test o n se nsiti vit y 
is 0. 0 4 7 5 a n d ac hie ve d b y t he test o n s pecificit y is 0. 0 4 1.  T he c urre nt recall rate (i.e., 
mar gi n p ositi vit y rate) after t he use of ca vit y s ha v e mar gi n is ~ 1 9 % 
3 6, w hic h a grees wit h 
t he best practices i n Was hi n gt o n U ni versit y, ~ 1 5. 5 %. 8 8 patie nts all o ws 9 2. 6 6 % p o wer t o 
test a re d ucti o n i n p ositi ve mar gi n rate fr o m 1 5. 5 % t o 5 % base d o n a o ne- si de d Bi n o mial 
e xact test at a 5 % le vel.   
 
 
8. 5  
E arl y st o p pi n g r ule 
 
We d o n’t e x pect a n y t o xicities f or L S 3 0 1 b ut gra de 1 t o xicit y is us uall y acce pta ble f or 
i ma gi n g a ge nt. As s u c h, t he trial will st o p earl y i n t he p hase 1 part f oll o wi n g t he r olli n g si x 
desi g n r ule if d ose le v el 1 has t w o o ut of si x patie nts e x perie nce D L Ts.  
 
8. 6  
A n al ysis set  
 
T he safet y a nal ysis set i ncl u des all patie nts w h o recei ve a n y a m o u nt of L S 3 0 1.  
 
T he effi cac y a nal ysis set i ncl u de all patie nts w h o recei ve a n y a m o u nt of L S 3 0 1, c o m plete 
t he s ur ger y a n d ha ve e val ua ble t u m or hist ol o g y res ults. 
 
 
 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 4 1  of 5 4  
 
 
 
9. 0  
R E F E R E N C E S  
 
1.  
C he n, S. L., I d di n gs, D. M., Sc heri, R. P. & Bilc hi k, A.J. L y m p hatic ma p pi n g a n d se nti nel 
n o de a nal ysis: c urr e nt c o nce pts a n d a p plicati o ns. C A C a ncer J. Cli n.  5 6 , 2 9 2- 3 0 9 ( 2 0 0 6). 
2.  
M ora n, M. S. , et al. S ociet y of S ur gical O nc ol o g y – A merica n S ociet y f or Ra diati o n 
O nc ol o g y C o nse ns us G ui deli ne o n Mar gi ns f or Breast- C o nser vi n g S ur ger y Wit h W h ole-
Breast Irr a diati o n i n Sta ges I a n d II I n vasi ve Breast Ca ncer. J o ur n al of Cli nic al O nc ol o gy  
( 2 0 1 4). 
3.  
Jac o bs, L. P ositi ve mar gi ns: T he c halle n ge c o nti n ues f or breast s ur ge o ns. A n n. S ur g. 
O nc ol.  1 5 , 1 2 7 1- 1 2 7 2 ( 2 0 0 8). 
4.  
C olli ns, L. , et al. O utc o me of W o me n wit h D uctal Carci n o ma I n Sit u ( D CI S) Treat e d 
wit h Breast - C o nser vi n g S ur ger y Al o ne: A Case- C o ntr ol St u d y of 2 2 5 Patie nts fr o m t he 
Ca ncer Res ear c h Net w or k. M o d. P at h ol.  2 2 , 3 4- 3 5 ( 2 0 0 9). 
5.  
Vici ni, F. A. , et al. I m pact of y o u n g a ge o n o utc o me i n patie nts wit h d uctal carci n o ma-i n-
sit u treate d wit h breast- c o nser vi n g t hera p y. J o ur n al of cli nic al o nc ol o gy : offici al j o ur n al 
of t he A meric a n S ociet y of Cli nic al O nc ol o gy  1 8 , 2 9 6- 3 0 6 ( 2 0 0 0). 
6.  
Mc Ca hill, L. E. , et al. Varia bilit y i n ree xcisi o n f oll o wi n g breast c o ns er vati o n s ur ger y. 
J a m a 3 0 7 , 4 6 7- 4 7 5 ( 2 0 1 2). 
7.  
M orr o w, M. , et al. S ur ge o n rec o m me n dati o ns a n d recei pt of m astect o m y f or treat me nt of 
breast ca ncer. J a m a 3 0 2 , 1 5 5 1- 1 5 5 6 ( 2 0 0 9). 
8.  
M ulle n, R. , et al. I n v ol ve d a nteri or mar gi ns after breast c o nser vi n g s ur ger y: Is re-e xcisi o n 
re q uire d ? E ur o pe a n J o ur n al of S ur gic al O nc ol o gy  3 8 , 3 0 2- 3 0 6. 
9.  
S kri pe n o va, S. & La yfiel d, L.J. I nitial mar gi n stat us f or i n vasi ve d uctal carci n o ma of t he 
breast a n d s u bse q u e nt i de ntificati o n of carci n o ma i n ree xcisi o n s peci me ns. Arc hives of 
p at h ol o gy & l a b or at ory me dici ne  1 3 4 , 1 0 9-1 1 4 ( 2 0 1 0).  
1 0.  
Jac o bs, L. P ositi ve mar gi ns: T he c halle n ge c o nti n ues f or breast s ur ge o ns. A n n als of 
S ur gic al O nc ol o gy  1 5 , 1 2 7 1- 1 2 7 2 ( 2 0 0 8). 
1 1.  
Za va g n o, G. , et al. R ole of resecti o n mar gi ns i n patie nts treate d wit h breast c o nser vati o n 
s ur ger y. C a ncer  1 1 2 , 1 9 2 3- 1 9 3 1 ( 2 0 0 8). 
1 2.  
Gi ulia n o, A. E., Kir ga n, D. M., G ue nt her, J. M. & M ort o n, D. L. L y m p hatic ma p pi n g a n d 
se nti nel l y m p ha de nect o m y f or breast ca ncer. A n n als of s ur gery  2 2 0 , 3 9 1- 3 9 8; disc ussi o n 
3 9 8- 4 0 1 ( 1 9 9 4). 
1 3.  
C o x, C. E. , et al. G ui deli nes f or se nti nel n o de bi o ps y a n d l y m p hatic ma p pi n g of patie nts 
wit h breast ca n cer. A n n als of s ur gery  2 2 7 , 6 4 5- 6 5 1; disc ussi o n 6 5 1- 6 4 3 ( 1 9 9 8). 
1 4.  
Nie we g, O. E. & V ee nstr a, H.J. False- ne g ati ve se nti nel n o de bi o ps y i n mela n o ma. J. S ur g. 
O nc ol.  1 0 4 , 7 0 9- 7 1 0 ( 2 0 1 1). 
1 5. 
Bez u, C. , et al. A na p h ylactic res p o nse t o bl ue d y e d uri n g se nti nel l y m p h n o de bi o ps y. 
S ur g. O nc ol.  2 0 , 5 5- 5 9 ( 2 0 1 1). 
1 6.  
va n de n Ber g, N. S. , et al. M ulti m o dal S ur gical G ui da nce d uri n g Se nti nel N o de Bi o ps y 
f or Mela n o ma: C o m bi ne d Ga m ma Traci n g a n d Fl u oresce n ce I m a gi n g of t h e Se nti nel 
N o de t hr o u g h Use of t he H y bri d Tracer I n d oc ya ni ne Gree n- Tc - Na n o c oll oi d. R a di ol o gy , 
1 4 0 3 2 2 ( 2 0 1 4). 
1 7. 
Arii, S. , et al. S ur gical strate gies f or he pat o cell ular carci n o ma wit h s pecial refere nce t o 
a nat o mical he patic resecti o n a n d i ntra o perati ve c o ntrast -e n ha n ce d ultras o n o gra p h y. 
O nc ol o gy  7 8 S u p pl 1 , 1 2 5- 1 3 0 ( 2 0 1 0). 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 4 2  of 5 4  
 
 
1 8.  
va n Vle d der, M. G. , et al. T he effect of steat osis o n ec h o ge nicit y of c ol orectal li ver 
metastases o n i ntra o perati ve ultras o n o gra p h y. Arc h S ur g  1 4 5 , 6 6 1- 6 6 7 ( 2 0 1 0). 
1 9.  
U ki m ura, O. , et al. I ntra o perati ve ultras o n o gra p h y i n a n era of mi ni mall y i n vasi ve 
ur ol o g y. I nter n ati o n al j o ur n al of ur ol o gy : offici al j o ur n al of t he J a p a nese Ur ol o gic al 
Ass oci ati o n  1 5 , 6 7 3- 6 8 0 ( 2 0 0 8). 
2 0.  
Ka ne, R. A. I ntr a o per ati ve ultras o n o gra p h y: hist or y, c urre nt state of t he art, a n d f ut ure 
directi o ns. J o ur n al of ultr as o u n d i n me dici ne : offici al j o ur n al of t he A meric a n I nstit ute 
of Ultr as o u n d i n Me dici ne  2 3 , 1 4 0 7- 1 4 2 0 ( 2 0 0 4). 
2 1.  
R u di n, M. & Weissle der, R. M olec ular i ma gi n g i n dr u g disc o ver y a n d de vel o p me nt. N at. 
Rev. Dr u g Disc o v.  2 , 1 2 3- 1 3 1 ( 2 0 0 3). 
2 2.  
Wil ke, L. G. , et al. Ra pi d n o ni n vasi ve o ptical i ma gi n g of tiss ue c o m p ositi o n i n breast 
t u m or mar gi ns. A meric a n J o ur n al of S ur gery  1 9 8 , 5 6 6- 5 7 4 ( 2 0 0 9). 
2 3.  
Tr o ya n, S. L. , et al. T he F L A R E(( T M)) I ntra o p erati ve Near -I nfrare d Fl u ores ce nce 
I ma gi n g S yste m: A First-i n-H u ma n Cli nical Trial i n Breast Ca ncer Se nti nel L y m p h N o de 
Ma p pi n g. A n n als of S ur gic al O nc ol o gy  1 6 , 2 9 4 3- 2 9 5 2 ( 2 0 0 9). 
2 4.  
Keller, M. D., Maj u m der, S. K. & Ma ha de va n- La nse n, A. S patiall y offset Ra ma n 
s pectr osc o p y of l a yer e d s oft tiss ues. O ptics Letters  3 4 , 9 2 6- 9 2 8 ( 2 0 0 9). 
2 5.  
B y dl o n, T. M. , et al. Ra pi d O ptical I ma gi n g of Breast T u m or Mar gi ns: Fi nal Res ults fr o m 
a 1 0 0-Patie nt Cli nical St u d y. C a ncer Rese ar c h  6 9 , 7 7 0 S- 7 7 1 S ( 2 0 0 9). 
2 6.  
B h us ha n, K. R. , et al. Detecti o n of Breast Ca n cer Micr ocalcifi cati o ns Usi n g a D ual -
m o dalit y S P E C T/ NI R Fl u oresce nt Pr o b e. J o ur n al of t he A meric a n C he mic al S ociety  1 3 0 , 
1 7 6 4 8- + ( 2 0 0 8). 
2 7.  
Marz ull o, A. C. D., Net o, O. P., Bitar, R. A., Marti n h o, H. D. & Marti n, A. A. F T- Ra ma n 
s pectra of t h e b or der of i nfiltrati n g d uctal carci n o ma lesi o ns. P h ot o me dici ne a n d L aser 
S ur gery  2 5 , 4 5 5- 4 6 0 ( 2 0 0 7). 
2 8.  
Z ys k, A. M. & B o p part, S. A. C o m p utati o nal met h o ds f or a nal ysis of h u ma n breast t u m or 
tiss ue i n o ptical c o here n ce t o m o gra p h y i ma ges. J o ur n al of Bi o me dic al O ptics  1 1 , - 
( 2 0 0 6). 
2 9.  
Ha ka, A. S. , et al. I n vi v o mar gi n assess me nt d uri n g partial mastect o m y breast s ur ger y 
usi n g Ra ma n s pectr osc o p y. C a ncer Rese ar c h  6 6 , 3 3 1 7- 3 3 2 2 ( 2 0 0 6). 
3 0.  
Le n ki ns ki, R. E., A h me d, M., Za heer, A., Fra n gi o ni, J. V. & G ol d ber g, S. N. Near- i nfrare d 
fl u oresce n ce i ma gi n g of micr ocalcificati o n i n a n a ni mal m o del of breast ca ncer. 
Ac a de mic R a di ol o gy  1 0 , 1 1 5 9- 1 1 6 4 ( 2 0 0 3). 
3 1.  
Z uza k, K.J., Sc hae berle, M. D., Gla d wi n, M. T., Ca n n o n, R. O. & Le vi n, I. W. N o ni n vasi ve 
deter mi nati o n of s patiall y res ol ve d a n d ti me-res ol ve d tiss ue perf usi o n i n h u ma ns d uri n g 
nitric o xi de i n hi biti o n a n d i n halati o n b y use of a visi ble- reflecta nce h y p ers pectral 
i ma gi n g tec h ni q ue. Circ ul ati o n  1 0 4 , 2 9 0 5- 2 9 1 0 ( 2 0 0 1). 
3 2.  
Tr o ya n, S. L. , et al. T he F L A R E I ntra o perati ve N ear -I nfrare d Fl u oresce nce I ma gi n g 
S yste m: A First -i n-H u ma n Cli nical Trial i n Breast Ca ncer Se nti nel L y m p h N o de 
Ma p pi n g. A n n. S ur g. O nc ol.  1 6 , 2 9 4 3- 2 9 5 2 ( 2 0 0 9). 
3 3. 
Hirc he, C. , et al. A n e x p eri me ntal st u d y t o e val uate t he Fl u o bea m 8 0 0 i ma gi n g s yste m 
f or fl u ores ce nce- g ui de d l y m p hatic i ma gi n g a n d se nti nel n o de bi o ps y. S ur gic al i n n ov ati o n  
2 0 , 5 1 6- 5 2 3 ( 2 0 1 3). 
3 4.  
T o bis, S. , et al. Near i nfr are d fl u oresce nce i ma gi n g after i ntra v e n o us i n d oc ya ni ne gree n: 
i nitial cli nical e x perie nce wit h o pe n partial ne p hrect o m y f or re n al c ortical t u m ors. 
Ur ol o gy  7 9 , 9 5 8- 9 6 4 ( 2 0 1 2). 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 4 3  of 5 4  
 
 
3 5.  
G ot o h, K. , et al. A n o vel i ma ge- g ui de d s ur ger y of he pat ocell ular carci n o ma b y 
i n d oc ya ni ne gree n fl u or esce nce i ma gi n g n a vi gati o n. J. S ur g. O nc ol.  1 0 0 , 7 5- 7 9 ( 2 0 0 9). 
3 6.  
Li u, Y. , et al. I ntr a o perati ve detecti o n of li ver t u m ors ai de d b y a fl u oresce n ce g o g gle 
s yste m a n d m ulti m o dal i ma gi n g. T he A n alyst  1 3 8 , 2 2 5 4- 2 2 5 7 ( 2 0 1 3). 
3 7.  
Li u, Y. , et al. Ha n ds-free, wireless g o g gles f or n ear-i nfrare d fl u oresce nce a n d real-ti me 
i ma ge- g ui de d s ur ger y. S ur gery  1 4 9 , 6 8 9- 6 9 8 ( 2 0 1 1). 
3 8.  
Li u, Y. , et al. Near -i nfrar e d fl u oresce nce g o g gle s yste m wit h c o m ple me ntar y metal-
o xi de-se mic o n d uct or i ma gi n g se ns or a n d see-t hr o u g h dis pla y. J o ur n al of bi o me dic al 
o ptics  1 8 , 1 0 1 3 0 3 ( 2 0 1 3). 
3 9.  
Li u, Y., S ol o m o n, M. & Ac hilef u, S. Pers pecti ves a n d p ote ntial a p plicati o ns of 
na n o me dici ne i n breast a n d pr ostate ca n cer. Me di ci n al rese arc h revie ws  3 3 , 3- 3 2 ( 2 0 1 3). 
4 0.  
Li u, Y. , et al. C o m ple me ntar y fl u ores ce nce- p olarizati o n micr osc o p y usi n g di visi o n- of-
f ocal- pla n e p olarizati o n i ma gi n g se ns or. J o ur n al of bi o me dic al o ptics  1 7 , 1 1 6 0 0 1 ( 2 0 1 2). 
4 1.  
Li u, Y. , et al. First i n - h u ma n i ntra o perati ve i ma gi n g of H C C usi n g t he fl u oresce n ce 
g o g gle s yste m a n d tra nsarterial deli ver y of near- i nfrare d fl u ores ce nt i ma gi n g a ge nt: a 
pil ot st u d y. Tr a nsl ati o n al rese arc h : t he j o ur n al of l a b or at ory a n d cli nic al me dici ne  1 6 2 , 
3 2 4- 3 3 1 ( 2 0 1 3). 
4 2.  
M o n dal, S. B. , et al. Real -ti me fl u oresce nce i ma g e- g ui de d o nc ol o gic s ur ger y. A dv a nces i n 
c a ncer res e arc h  1 2 4 , 1 7 1- 2 1 1 ( 2 0 1 4). 
4 3.  
M o n dal, S. B. , et al. Bi n oc ular G o g gle A u g me nte d I ma gi n g a n d Na vi gati o n S yste m 
pr o vi des real-ti me fl u oresce nce i ma g e g ui da nce f or t u m or resecti o n a n d se nti nel l y m p h 
n o de ma p pi n g. Sci Re p  5 , 1 2 1 1 7 ( 2 0 1 5). 
4 4. 
Kra mer - Mar e k, G., G or e, J. & K orc, M. M olec ular i ma gi n g i n pa ncreatic ca ncer--a 
r oa d ma p f or t h era p e utic decisi o ns. C a ncer letters  3 4 1 , 1 3 2- 1 3 8 ( 2 0 1 3). 
4 5.  
Ya n g, L., Ca o, Z., Li n, Y., W o o d, W. C. & Stale y, C. A. M olec ular beac o n i ma gi n g of 
t u m or mar ker ge n e e x pressi o n i n pa ncreatic ca ncer cells. C a ncer bi ol o gy & t her a py  4 , 
5 6 1- 5 7 0 ( 2 0 0 5). 
4 6. 
Bares, R. , et al. F- 1 8 fl u or o de o x y gl uc ose P E T i n vi v o e val uati o n of pa ncreatic gl uc ose 
meta b olis m f or detecti o n of pa ncreatic ca ncer. R a di ol o gy  1 9 2 , 7 9- 8 6 ( 1 9 9 4). 
4 7.  
Traj k o vic- Arsic, M. , et al. M ulti m o dal m olec ular i ma gi n g of i nte gri n al p ha v beta 3 f or i n 
vi v o detecti o n of pa ncreatic ca ncer. J o ur n al of n ucle ar me dici ne : offici al p u blic ati o n, 
S ociety of N ucle ar Me di ci ne  5 5 , 4 4 6- 4 5 1 ( 2 0 1 4). 
4 8.  
Carls o n, S. K. , et al. Q ua ntitati ve m olec ular i ma gi n g of viral t hera p y f or pa ncreatic ca n cer 
usi n g a n e n gi neer e d measles vir us e x pressi n g t he s o di u m-i o di de s y m p orter re p orter g e ne. 
AJ R. A meric a n j o ur n al of r oe nt ge n ol o gy  1 9 2 , 2 7 9- 2 8 7 ( 2 0 0 9). 
4 9. 
Eser, S. , et al. I n vi v o dia g n osis of m uri ne pa ncreatic i ntrae pit helial ne o plasia a n d earl y-
sta ge pa ncreatic ca ncer b y m olec ular i ma gi n g. Pr oc N atl Ac a d Sci U S A  1 0 8 , 9 9 4 5- 9 9 5 0 
( 2 0 1 1). 
5 0. 
He, Y. , et al. A nti - C X C R 4 m o n ocl o nal a nti b o d y c o nj u gate d t o ultras mall 
s u per par a ma g netic ir o n o xi de na n o particles i n a n a p plicati o n of M R m olec ular i ma gi n g 
of pa ncr eatic ca ncer cell li nes. Act a r a di ol o gic a  5 3 , 1 0 4 9- 1 0 5 8 ( 2 0 1 2). 
5 1.  
Ya n g, L. , et al. M olec ular i ma gi n g of pa ncreatic ca ncer i n a n a ni mal m o del usi n g 
tar gete d m ultif u ncti o nal na n o particles. G astr oe nt er ol o gy  1 3 6 , 1 5 1 4- 1 5 2 5 e 1 5 1 2 ( 2 0 0 9). 
5 2.  
Keli her, E.J. , et al. Tar geti n g cat he psi n E i n pa ncreatic ca ncer b y a s mall m olec ule all o ws 
i n vi v o detecti o n. Ne o pl asi a  1 5 , 6 8 4- 6 9 3 ( 2 0 1 3). 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 4 4  of 5 4  
 
 
5 3. 
Li u, Z. , et al. I nte gri n al p ha v beta 6-T ar get e d S P E C T I m a gi n g f or Pa ncreatic Ca ncer 
Detecti o n. J o ur n al of n ucle ar me dici ne : offici al p u blic ati o n, S ociety of N ucle ar 
Me dici ne  5 5 , 9 8 9-9 9 4 ( 2 0 1 4).  
5 4.  
Bre mer, C., T u n g, C. H. & Weissle der, R. M olec ular i ma gi n g of M M P e x pressi o n a n d 
t hera pe utic M M P i n hi biti o n. Ac a de mic r a di ol o gy  9 S u p pl 2 , S 3 1 4- 3 1 5 ( 2 0 0 2). 
5 5. 
Lee, H. , et al. N ear -i nfrar e d p H-acti vata ble fl u ores ce nt pr o bes f or i ma gi n g pri mar y a n d 
metastatic breast t u m ors. Bi oc o nj u g ate c he mistry  2 2 , 7 7 7- 7 8 4 ( 2 0 1 1). 
5 6.  
Ura n o, Y. , et al. Selecti ve m olec ular i ma gi n g of via ble ca n cer cells wit h p H -acti vata bl e 
fl u oresce n ce pr o bes. N at ure me dici ne  1 5 , 1 0 4- 1 0 9 ( 2 0 0 9). 
5 7.  
Ac hilef u, S., D ors h o w, R. B., B u gaj, J. E. & Raja g o pala n, R. N o vel rece pt or- tar gete d 
fl u oresce nt c o ntrast a g e nts f or i n vi v o t u m or i ma gi n g. I nvesti g ative r a di ol o gy 3 5 , 4 7 9-
4 8 5 ( 2 0 0 0). 
5 8.  
B u gaj, J. E., Ac hilef u, S., D ors h o w, R. B. & Raja g o pala n, R. N o vel fl u ores ce nt c o ntrast 
a ge nts f or o ptical i ma gi n g of i n vi v o t u m ors base d o n a rece pt or-tar gete d d ye- pe pti de 
c o nj u gate platf or m. J o ur n al of bi o me dic al o ptics  6 , 1 2 2- 1 3 3 ( 2 0 0 1). 
5 9.  
Ac hilef u, S., Wil hel m, R. R., Ji me nez, H. N., Sc h mi dt, M. A. & Sri ni vasa n, A. A ne w 
met h o d f or t he s y nt hesis of tri- tert-b ut yl diet h yle n etria mi ne pe ntaacetic aci d a n d its 
deri vati ves. T he J o ur n al of or g a nic c he mistry  6 5 , 1 5 6 2- 1 5 6 5 ( 2 0 0 0). 
6 0.  
Ac hilef u, S. , et al. S y ner gistic effects of li g ht-e mitti n g pr o bes a n d pe pti des f or tar geti n g 
a n d m o nit ori n g i nte gri n e x pressi o n. Pr oc. N atl. Ac a d. Sci. U. S. A.  1 0 2 , 7 9 7 6- 7 9 8 1 
( 2 0 0 5). 
6 1.  
Ac hilef u, S. Li g hti n g u p t u m ors wit h rece pt or-s p ecific o ptical m olec ular pr o bes. 
Tec h n ol o gy i n C a ncer Rese arc h & Tre at me nt  3 , 3 9 3- 4 0 9 ( 2 0 0 4). 
6 2.  
Ac hilef u, S., D ors h o w, R. B., B u gaj, J. E. & Raja g o pala n, R. N o vel rece pt or- tar gete d 
fl u oresce nt c o ntrast a g e nts f or i n vi v o t u m or i ma gi n g. I nvesti g ative R a di ol o gy 3 5 , 4 7 9-
4 8 5 ( 2 0 0 0). 
6 3. 
Berezi n, M. Y. , et al. Rati o nal a p pr oac h t o select s mall pe pti de m olec ular pr o bes la bele d 
wit h fl u oresce nt c y a ni ne d yes f or i n vi v o o ptical i ma gi n g. Bi oc he mistry  5 0 , 2 6 9 1- 2 7 0 0 
( 2 0 1 1). 
6 4.  
G oiff o n, R.J., A kers, W.J., Berezi n, M. Y., L ee, H. & Ac hilef u, S. D y na mic n o ni n vasi ve 
m o nit ori n g of re nal f u ncti o n i n vi v o b y fl u oresce n ce lifeti me i ma gi n g. J o ur n al of 
bi o me dic al o ptics  1 4 , 0 2 0 5 0 1 ( 2 0 0 9). 
6 5. 
T h ota, R., Pa uff, J. M. & Berli n, J. D. Treat me nt of metastatic pa ncreatic a d e n ocarci n o ma: 
a re vie w. O nc ol o gy  2 8 , 7 0- 7 4 ( 2 0 1 4). 
6 6. 
Al - Hajeili, M., Az mi, A. S. & C h oi, M. Na b-paclita xel: p ote ntial f or t he treat me nt of 
a d va nce d pa ncreatic ca n cer. O nc o T ar gets a n d t her a py  7 , 1 8 7- 1 9 2 ( 2 0 1 4). 
6 7.  
Kratz, F. Al b u mi n as a dr u g carri er: desi g n of pr o dr u gs, dr u g c o nj u gates a n d 
na n o particles. J o ur n al of c o ntr olle d rele ase : offici al j o ur n al of t he C o ntr olle d Rele ase 
S ociety  1 3 2 , 1 7 1- 1 8 3 ( 2 0 0 8). 
6 8.  
C hara n ya, T. , et al. Tri m o dal c ol or-fl u oresce n ce- p olarizati o n e n d osc o p y ai de d b y a t u m or 
selecti ve m olec ular pr o b e acc uratel y d etects flat l esi o ns i n c olitis-ass ociate d ca ncer. 
J o ur n al of bi o me dic al o ptics  1 9 , 1 2 6 0 0 2 ( 2 0 1 4). 
6 9.  
R ose nt hal, E. L. , et al. Safet y a n d T u m or S pecificit y of Cet u xi ma b-I R D ye 8 0 0 f or S ur gical 
Na vi gati o n i n Hea d a n d Nec k Ca ncer. Cli nic al C a ncer Rese ar c h  2 1 , 3 6 5 8- 3 6 6 6 ( 2 0 1 5). 
7 0.  
Zi n n, K. R. , et al. I N D- Directe d Safet y a n d Bi o distri b uti o n St u d y of I ntra v e n o usl y 
I njecte d Cet u xi ma b-I R D ye 8 0 0 i n C y n o m ol g us Maca q u es. M olec ul ar i m a gi n g a n d 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  0 5/ 1 5/ 2 0 2 3 ( A3)  
 
Pa ge 4 5  of 5 4  
 
 
bi ol o gy : MI B : t he offici al p u blic ati o n of t he Ac a de my of M olec ul ar I m a gi n g  1 7 , 4 9- 5 7 
( 2 0 1 5). 
7 1.  
Ga y ke ma, S. B. M. , et al. 8 9 Zr-Be vaciz u ma b P E T I ma gi n g i n Pri mar y Breast Ca ncer. 
J o ur n al of N ucle ar Me di ci ne  5 4 , 1 0 1 4- 1 0 1 8 ( 2 0 1 3). 
7 2.  
Fra n kli n, H. L., Miller, D. M., He d ges, T., Perr y, J. & Parris h- N o va k, J. Cli nical 
de vel o p me nt of B L Z- 1 0 0 f or real -ti me o ptical i ma gi n g of t u m ors d uri n g res ecti o n. V ol. 
9 6 9 6 9 6 9 6 0 V- 9 6 9 6 0 V- 9 6 9 6 7 ( 2 0 1 6). 
7 3.  
Fi del, J. , et al. Precli nical Vali dati o n of t he Utilit y of B L Z - 1 0 0 i n Pr o vi di n g Fl u oresce nce 
C o ntrast f or I ma gi n g S p o nta ne o us S oli d T u m ors. C a ncer Rese ar c h  7 5 , 4 2 8 3- 4 2 9 1 
( 2 0 1 5). 
7 4.   
S k ol ni k J M, Barrett J S, Ja yara m a n B, Patel D, A d a ms o n P C ( 2 0 0 8) S h orte ni n g t he 
ti meli ne of pe diatric p has e I trials: t he r olli n g si x desi g n. J Cli n O nc ol  2 6 , 1 9 0- 1 9 5. 
7 5.  
D o ussa u A, Ge o er ger B, Ji me nez I, Pa oletti X. I n n o vati o ns f or p hase I d ose-fi n di n g 
desi g ns i n pe diatric o nc ol o g y cli nical trials. C o nte m p orar y cli nical trials. 2 0 1 6. 4 7: 2 1 7-
2 2 7 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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1 0. 0  
A P P E N DI X A:  D efi niti o ns f or A d verse E ve nt Re p orti n g  
 
A.  A d verse E ve nts ( A Es)  
 
As defi ne d i n 2 1 C F R 3 1 2. 3 2: 
 
Defi niti o n:  a n y u nt o war d me dical occ urre n ce ass ociate d wit h t he use of a dr u g i n h u ma ns, 
w het her or n ot c o nsi dere d dr u g- relat e d. 
 
Gr a di n g:  t he descri pti o ns a n d gra di n g scales f o u n d i n t he re vise d N CI C o m m o n Ter mi n ol o g y 
Criteria f or A d verse E ve nts ( C T C A E) versi o n 5. 0 will be utilize d f or all t o xicit y re p orti n g.  A 
c o p y of t he C T C A E versi o n 5. 0 ca n be d o w nl o a de d fr o m t he C T E P we bsite. 
 
Attri b uti o n (rel ate d ness), E x pecte d ness, a n d Seri o us ness:  t he defi niti o ns f or t he ter ms 
liste d t hat s h o ul d be use d are t h ose pr o vi de d b y t he De part me nt of Healt h a n d H u ma n Ser vices’ 
Office f or H u ma n Researc h Pr otecti o ns ( O H R P).  A c o p y of t his g ui da n ce ca n b e f o u n d o n 
O H R P’s we bsite:  
htt p:// w w w. h hs. g o v/ o hr p/ p olic y/a d ve v nt g ui d. ht ml  
 
B.  S us pecte d A d verse Re a cti o n ( S A R)  
 
As defi ne d i n 2 1 C F R 3 1 2. 3 2: 
 
Defi niti o n:  a n y a d verse e ve nt f or w hic h t here is a reas o na ble p ossi bilit y t hat t he dr u g ca use d 
t he a d verse e ve nt.  “ R eas o na ble p ossi bilit y” mea ns t here is e vi de nce t o s u g gest a ca usal 
relati o ns hi p bet wee n t he dr u g a n d t he a d vers e e ve nt.  “ S us pecte d a d v erse reacti o n” i m plies a 
lesser de gree of certai nt y a b o ut ca usalit y t ha n a d verse reacti o n, w hic h mea ns a n y a d verse e ve nt 
ca use d b y a dr u g. 
 
C.  Life -T hre ate ni n g A d verse E ve nt / Life T hre ate ni n g S us pecte d A d verse Re acti o n  
 
As defi ne d i n 2 1 C F R 3 1 2. 3 2: 
 
Defi niti o n:  a n y  a d v ers e  dr u g  e v e nt  or  s us p ecte d  a d vers e  reacti o n  is  c o nsi dere d  “life -
t hreate ni n g” if, i n t he vie w of t he i n vesti gat or, its occ urre nce places t he patie nt at i m me diate 
ris k of deat h. It d o es n ot i ncl u de a n a d v erse e ve nt or s us pect e d a d v erse reacti o n t hat, ha d it 
occ urr e d i n a m ore s e ver e f or m, mi g ht ha ve ca use d deat h. 
 
D.   Seri o us A d verse E ve nt ( S A E) or Seri o us S us pecte d A d vers e Re a cti o n  
 
As defi ne d i n 2 1 C F R 3 1 2. 3 2: 
 
Defi niti o n:  a n a d verse e ve nt or s us pecte d a d verse reacti o n is c o nsi dere d “seri o us” if, i n t he 
vie w of t he i n vesti gat or, it res ults i n a n y of t he f oll o wi n g o utc o mes: 

 
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Pa ge 4 7  of 5 4  
 
 
o  Deat h  
o  A life -t hreate ni n g a d vers e e ve nt 
o  I n p atie nt h os pitalizati o n or pr ol o n gati o n of e xisti n g h os pitalizati o n 
o  A persiste nt or si g nifica nt i nca pacit y or s u bsta ntial disr u pti o n of t he a bilit y t o c o n d uct 
n or mal life f u ncti o ns 
o  A c o n ge nital a n o mal y/ birt h defect 
o  A n y ot her i m p orta nt me dical e ve nt t hat d oes n ot fit t he criteria a b o ve b ut, b ase d u p o n 
a p pr o priate me dical j u d g me nt, ma y je o par dize t he s u bject a n d ma y re q uire me dical or 
s ur gical i nter ve nti o n t o pre ve nt o ne of t he o utc o m es liste d a b o ve 
 
E.  Pr ot oc ol E xce pti o ns 
 
Defi niti o n:  A pla n ne d c h a n ge i n t he c o n d u ct of t he researc h f or o ne p artici pa nt. 
 
F.  De vi ati o n 
 
Defi niti o n:  A n y alterati o n or m o dificati o n t o t h e I R B -a p pr o ve d researc h wit h o ut pr os pecti ve 
I R B a p pr o val.  T h e ter m “researc h ” e n c o m passes all I R B-a p pr o ve d m ateri als a n d d oc u me nts 
i ncl u di n g  t he  detaile d  pr ot oc ol,  I R B  a p plicati o n,  c o nse nt  f or m,  recr uit me nt  materials,  
q uesti o n naires/ data c ollecti o n f or ms, a n d a n y ot her i nf or mati o n relati n g t o t he researc h st u d y. 
 
A mi n or or a d mi nistrati ve de viati o n is o ne t hat d oes n ot ha ve t he p ote ntial t o ne gati vel y i m pact 
t he ri g hts, safet y, or w elf are of p artici pa nts or ot h ers or t he sci e ntific vali dit y of t he st u d y. 
 
A maj or de viati o n is o ne t hat d oes ha ve t he p ote ntial t o ne gati vel y i m pact t he ri g hts, safet y, or 
welfar e of partici pa nts or ot hers or t he scie ntific v ali dit y of t he st u d y. 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Pr ot oc ol Versi o n:  1 1/ 4/ 2 0 2 1 ( A 2) 
 
Pa ge 4 8  of 5 4  
1 1. 0  
A P P E N DI X B:  Re p orti n g Ti meli nes  
 
 
 
E x pe dite d Re p orti n g Ti meli nes  
E ve nt  
H R P O –  W as hi n gt o n 
U ni versit y  
H R P P O –  U T  
S o ut h wester n  
Q A S M C –  
W as hi n gt o n 
U ni versit y  
D S M C –  U T  
S o ut h wester n  
F D A  
Seri o us A N D u ne x pecte d 
s us pecte d a d verse reacti o n 
 
 
 
S u b mit S A E 
Re p ort t o S C C C 
D S M C  wit hi n 5 
b usi ness da ys of 
st u d y tea m 
a ware ness of t he 
e ve nt(s) 
Re p ort n o later t ha n 1 5 cale n dar 
da ys after it is deter mi ne d t hat t he 
i nf or mati o n q ualifies f or 
re p orti n g 
U ne x pecte d fatal or life -
t hreate ni n g s us pecte d a d verse 
reacti o n 
 
 
 
S u b mit wit hi n 5 
b usi ness da ys of 
st u d y tea m 
a ware ness of t he 
e ve nt(s) 
Re p ort n o later t ha n 7 cale n dar 
da ys after i nitial recei pt of t he 
i nf or mati o n 
U na ntici pate d pr o ble m 
i n v ol vi n g ris k t o partici pa nts or 
ot hers 
Re p ort wit hi n 1 0 
w or ki n g da ys.  If t he 
e ve nt res ults i n t he 
deat h of a partici pa nt 
e nr olle d at 
W U/ BJ H/ S L C H, 
re p ort wit hi n 1 
w or ki n g da y.  
S u b mit wit hi n 5 
b usi ness da ys of 
st u d y tea m a ware ness 
of t he e ve nt(s)  
Re p ort via e mail 
after I R B 
ac k n o wle d g me nt 
S u b mit U PI R S O 
Re p ort t o S C C C 
D S M C  wit hi n 5 
b usi ness da ys of 
st u d y tea m 
a ware ness of t he 
e ve nt(s) 
 
Maj or de viati o n  
Re p ort wit hi n 1 0 
w or ki n g da ys.  If t he 
e ve nt res ults i n t he 
deat h of a partici pa nt 
e nr olle d at 
W U/ BJ H/ S L C H, 
re p ort wit hi n 1 
w or ki n g da y.  
S u b mit wit hi n 5 
b usi ness da ys of 
st u d y tea m a ware ness 
of t he e ve nt(s)  
 
 
Re p ort n o later t ha n 1 5 cale n dar 
da ys after it is deter mi ne d t hat t he 
i nf or mati o n q ualifies f or 
re p orti n g 

 
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Pa ge 4 9  of 5 4  
 
 
 
 
E x pe dite d Re p orti n g Ti meli nes  
E ve nt  
H R P O –  W as hi n gt o n 
U ni versit y  
H R P P O –  U T  
S o ut h wester n  
Q A S M C –  
W as hi n gt o n 
U ni versit y  
D S M C –  U T  
S o ut h wester n  
F D A  
A series of mi n or de viati o ns 
t hat are bei n g re p orte d as a 
c o nti n ui n g n o nc o m plia nce 
Re p ort wit hi n 1 0 
w or ki n g da ys.   
S u b mit wit hi n 5 
b usi ness da ys of 
st u d y tea m a ware ness 
of t he e ve nt(s)  
 
 
 
Pr ot oc ol e xce pti o n  
A p pr o val m ust be 
o btai ne d pri or t o 
i m ple me nti n g t he 
c ha n ge 
A p pr o val m ust be 
o btai ne d pri or t o 
i m ple me nti n g t he 
c ha n ge 
 
 
 
Cli nicall y i m p orta nt i ncrease i n 
t he rate of a seri o us s us pecte d 
a d verse reacti o n of t hat list i n 
t he pr ot oc ol or I B 
 
 
 
 
Re p ort n o later t ha n 1 5 cale n dar 
da ys after it is deter mi ne d t hat t he 
i nf or mati o n q ualifies f or 
re p orti n g 
C o m plai nts  
If t he c o m plai nt 
re veals a n 
u na ntici pate d pr o ble m 
i n v ol vi n g ris ks t o 
partici pa nts or ot hers 
O R n o nc o m plia nce, 
re p ort wit hi n 1 0 
w or ki n g da ys.  If t he 
e ve nt res ults i n t he 
deat h of a partici pa nt 
e nr olle d at 
W U/ BJ H/ S L C H, 
re p ort wit hi n 1 
w or ki n g da y.  
Ot her wis e, re p ort at 
t he ti me of c o nti n ui n g 
re vie w. 
S u b mit wit hi n 5 
b usi ness da ys of 
st u d y tea m a ware ness 
of t he e ve nt(s)  
 
 
 
Breac h of c o nfi de ntialit y  
Wit hi n 1 0 w or ki n g 
da ys.  
 
 
 
 

 
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Pa ge 5 0  of 5 4  
 
 
 
 
E x pe dite d Re p orti n g Ti meli nes  
E ve nt  
H R P O –  W as hi n gt o n 
U ni versit y  
H R P P O –  U T  
S o ut h wester n  
Q A S M C –  
W as hi n gt o n 
U ni versit y  
D S M C –  U T  
S o ut h wester n  
F D A  
I ncarcerati o n 
If wit h dra wi n g t he 
partici pa nt p oses a 
safet y iss ue, re p ort 
wit hi n 1 0 w or ki n g 
da ys.   
 
If wit h dra wi n g t he 
partici pa nt d oes n ot 
re prese nt a safet y 
iss ue a n d t he patie nt 
will be wit h dra w n, 
re p ort at c o nti n ui n g 
re vie w. 
 
 
 
 
 
 
 
R o uti ne Re p orti n g Ti meli nes  
E ve nt  
H R P O –  W as hi n gt o n 
U ni versit y  
H R P P O –  U T 
S o ut h wester n  
Q A S M C –  
W as hi n gt o n 
U ni versit y  
D S M C –  U T  
S o ut h wester n  
F D A  
A d verse e ve nt or S A E t hat d oes 
n ot re q uire e x pe dite d re p orti n g  
If t he y d o n ot meet t he 
defi niti o n of a n 
u na ntici pate d pr o ble m 
i n v ol vi n g ris ks t o 
partici pa nts or ot hers, 
re p ort s u m mar y 
i nf or mati o n at t he 
ti me of c o nti n ui n g 
re vie w 
A d verse e ve nts will 
be re p orte d at 
C o nti n ui n g Re vie w.  
A d verse e ve nts will 
be re p orte d i n t he 
t o xicit y ta ble i n t he 
D S M re p ort w hic h is 
t y picall y d ue e ver y 6 
m o nt hs. 
A d verse eve nt s 
will be re vie we d 
at A n n ual D S M C 
Re vie w  
T he m ost c urre nt t o xicit y ta ble 
fr o m t he D S M re p ort is pr o vi de d 
t o t he F D A wit h t he I N D’s a n n ual 
re p ort. 
Mi n or de viati o n  
Re p ort s u m mar y 
i nf or mati o n at t he 
ti me of c o nti n ui n g 
re vie w. 
Mi n or de viati o ns will 
be re p orte d at 
C o nti n ui n g Re vie w. 
 
Mi n or d e viati o ns 
will be  re vie we d 
at A n n ual D S M C 
Re vie w  
 

 
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Pa ge 5 1  of 5 4  
 
 
C o m plai nts  
If t he c o m plai nt 
re veals a n 
u na ntici pate d pr o ble m 
i n v ol vi n g ris ks t o 
partici pa nts or ot hers 
O R n o nc o m plia nce, 
re p ort wit hi n 1 0 
w or ki n g da ys.  If t he 
e ve nt res ults i n t he 
deat h of a partici pa nt 
e nr olle d at 
W U/ BJ H/ S L C H, 
re p ort wit hi n 1 
w or ki n g da y.  
Ot her wis e, re p ort at 
t he ti me of c o nti n ui n g 
re vie w. 
All c o m plai nts are 
s u m marize d i n t he 
ne xt pr o gress re p ort 
s u b mitte d as part of 
c o nti n uati o n re vie w 
or i n t he fi nal 
re p ort s u b mitte d t o 
i nacti vate t he st u d y 
 
 
 
I ncarcerati o n 
 
 
 
 
 
 
 
If wit h dra wi n g t he 
partici pa nt p oses a 
safet y iss ue, re p ort 
wit hi n 1 0 w or ki n g 
da ys.   
 
If wit h dra wi n g t he 
partici pa nt d oes n ot 
re prese nt a safet y 
iss ue a n d t he patie nt 
will be wit h dra w n, 
re p ort at c o nti n ui n g 
re vie w. 
 
 
 
 

 
S T U 2 0 2 2- 0 3 8 5, Farr, F or m A 3 -Pr ot oc ol, M o d _ 3, 0 6 - 0 6-2 3 ( 2)  
 
Pr ot oc ol Versi o n:  1 1/ 4/ 2 0 2 1 ( A 2) 
 
Pa ge 5 2  of 5 4  
1 2. 0  
A P P E N DI X C:  W as hi n gt o n U ni versit y U n a ntici p ate d Pr o ble m Re p orti n g C o ver 
S heet  
 
S A E C O V E R S H E E T - Sec o n d ar y Site Assess me nt  
 
Was hi n gt o n U ni versit y H R P O #:  
S p o ns or -I n v esti gat or:  
S u bject I nitials:   
S u bject I D:  
Treati n g M D:  
Treati n g Site:  
E V E N T T E R M:  
A d missi o n Date:  
E V E N T G R A D E:  
Date of site’s first n otificati o n:  
 
Tre ati n g M D E ve nt Ass ess m e nt:  
 
 
Is t his e v e nt p ossi bl y, pr o b a bl y, or defi nitel y  relate d st u d y treat me nt? 
 
 yes    
 
 n o 
 
If yes, pleas e list w hic h dr u g (if m ore t ha n o n e) _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
       
E x pl ai n _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _           _ _ _ _ _ _ _ _ _ _ _ _ _ 
 
 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _                            _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _  
           _ _ _ _ _ _ _ 
P h ysici a n’s N a m e  
  
 
P h ysici a n’s Si g n at ure   
 
D ate  
 
 
 
 
 
 
 
 
 
 
 
 
 

 
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Pa ge 5 4  of 5 4  
Re p orter Na m e:  
                        Pl e as e Pri nt  
D at e
: 
 
I n v esti gat or’s N a me: 
                        Pl e as e Pri nt  
I n v esti gat or Si g nat ure: 
 
D at e
: 
            /                    / 
I n v esti gat or’s N a me: 
_ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _
_ _ 
                        Pl e as e Pri nt  
F or M ulti- Ce nter St u dies O nl y 
I n v esti gat or Si g nat ure: 
 
D at e
: 
            /                    / 
Sc a n a n d e m ail t he c o m plete d re p ort wit h s o urce d oc u me nt ati o n (if av ail a ble) t o:  
S C C D S M C @ U T S o ut h w e st er n. e d u  f or re p orti n g t o t he D S M C ( D at a S afety a n d 
M o nit ori n g C o m mittee)