Supplemental Figure Legend Supplemental Figure 1 A · Supplemental Figure Legend Supplemental Figure 1 (A) Principal component analysis of gene expression of Panc-1 and BxPC3 cells.All

Supplemental Figure Legend

Supplemental Figure 1

(A) Principal component analysis of gene expression of Panc-1 and BxPC3 cells. All

controls: [Ctrl: untreated cells, Ctrl Exo: control exosomes (no siRNA payload), Scr

(siScrbl Exo), sirnaExo (exosomes with KrasG12D siRNA payload)]. (B-E) Heat map

depicting expression of top 1,000 genes (B-D) and differentially expressed genes

(statistically significant) (C-E) in Panc-1 (B, C) and BxPC3 (D, E) cells. Red: sirnaExo:

exosomes with KrasG12D siRNA payload, black: Scr: siScrl Exo, dark grey: CtrlExo: control

exosomes (no siRNA payload), light grey: Ctrl (untreated cells). (F) Gene set enrichment

plot for significantly down and up-regulated genes in siKrasG12D-1 Exo treated Panc-1 cells

compared to all controls.


(A) Sanger sequencing of PDX tumors (n=2 mice), to confirm the G to A base substitution

associated with the glycine to aspartic acid mutation at codon 12 (G12D). (B) Comparison

of quantification of tumor burden pre-treatment (baseline) by ultrasound (US, day 58 post

tumor induction), and MRI (day 60 post tumor induction), siKrasG12D-1 iExo (n=7) and

siScrbl iExo (n=7). Each set of bars for US and MRI represents measures for individual

mice. (C) Detailed findings from individual PDX mouse treated with siKrasG12D-1 iExo (n=7)

and siScrbl iExo (n=7), showing survival post treatment start (PTS) and post birth (PB),

baseline pre-treatment tumor volumes as measure by both ultrasound (US) and MRI,

post-treatment MRI measurements of tumor volume, tumor weight (TW) and body weight

(BW) at end point, and observations at necropsy/euthanasia. No measurements for MRI

was recorded if the mice were euthanized or the imaging was of poor quality (CNBM:

could not be measured). N/A: not applicable. (D) Pictures of H&E stained tissue sections

showing lesions of inflammation observed in organs of a siKrasG12D-1 iExo treated mouse

(A1-B). See accompanying source data.


(A) Number of exosomes per cell per hour determine by NanoSightTM for each MSC

donors listed. Donor 3 was used for all subsequent MSCs-derived exosomes studies.

Three independent measurements from distinct conditioned media collection are shown.

(B-C) Flow cytometry analyses histogram plots of the listed cell surface markers for BJ

fibroblast cells (B) and mesenchymal stem cells, MSCs (C). The inset numbers depict the

percent positive cells. In grey: isotype control. See accompanying source data.


(A) Schematic detailing the procedure for MSCs bioreactor culture for collection of

exosomes (B) Schematic of the isolation and electroporation procedures of exosomes

from MSCs conditioned media. (C) Schematic depicting the generation of MSCs-derived

iExosomes.


(A) Tabulated data depicting the quantitation of total exosomal protein and number of

exosomes in each of the 6 consecutive 48 hours harvests of 250 ml MSCs conditioned

media from the bioreactor. The data are also depicted in Figure 3A-C. (B) Quantitation of

the number of exosomes produced from three distinct bioreactors. (C) Quantitation of the

levels of glucose and lactose in the bioreactor, for the total duration of bioreactor

experiment (21 days). (D) Representative histogram of flow cytometry analysis of

exosomal markers (CD9, CD63, CD81) and CD47 on exosomes from all bioreactor

harvests (Bioreactor harvests 1 and 6 data are also shown in Figure 3E). Inset numbers

represent the percent positive beads. In grey: isotype control. See accompanying source

data.


(A) Quantification of apoptosis assayed by flow cytometry analyses in 689KPC cells (n=2

distinct experiments). One-way ANOVA comparing experimental groups to untreated. (B)

Images of H&E stained tissue sections showing representative lesions observed in the

pancreas, liver, spleen and kidney of KPC689 mice at experimental endpoint. Control Exo

(n=4), BJ siKrasG12D-1 iExo (n=6) and MSCs siKrasG12D-1 iExo (n=6). Scale bar: 100 μm.

(C) Percent mice with macroscopic metastases in the indicated organs at time of

necropsy. Control Exo (n=4), BJ siKrasG12D-1 iExo (n=6) and MSCs siKrasG12D-1 iExo

(n=6). (D) Detailed findings from individual mice with KPC689 tumors treated with Control

Exo (n=4), BJ siKrasG12D-1 iExo (n=6) and BJ siKrasG12D-1 iExo (n=6), showing survival

post treatment start (PTS) and post birth (PB), tumor weight (TW) and body weight (BW),

number of lung macrometastases, and observations at necropsy/euthanasia. (E) Tumor

burden (%) of KPC689 mice at experimental endpoint. Control Exo (n=4), BJ siKrasG12D-

1 iExo (n=6) and MSCs siKrasG12D-1 iExo (n=6). (F) Tumor weights (g: grams) of KPC689

mice at experimental endpoint. Control Exo (n=4), BJ siKrasG12D-1 iExo (n=6) and MSCs

siKrasG12D-1 iExo (n=6). (G) KPC689 orthotopic tumor growth (bioluminescence), and

average total flux at day 51, post tumor induction. Control Exo (n=4), BJ siKrasG12D-1 iExo

(n=6) and MSCs siKrasG12D-1 iExo (n=6). The mean +/- SEM is depicted and statistical

tests are listed for relevant panels. ** p<0.01. See accompanying source data.


(A) Detailed findings from individual mice with Panc-1 tumors treated with Control Exo

(n=4), BJ siKrasG12D-1 iExo (n=6) and BJ siKrasG12D-1 iExo (n=6), showing survival post

treatment start (PTS) and post birth (PB), tumor weight (TW) and body weight (BW), and

observations at necropsy/euthanasia. ** this tumor weight was not included; the pancreas

of this mouse was found grossly and histopathologically normal. (B) Pictures of H&E

stained sections of tumors from Panc-1 tumor bearing mice. Control Exo (n=5), MSCs

siKrasG12D-1 iExo, CB (n=5), MSCs siKrasG12D-1 iExo, RB (n=4), BJ siKrasG12D-1 iExo, RB

(n=5). Scale bars: 100 μm (top) and 50 μm (bottom). These data are also depicted in

Figure 4O. (C) Representative pictures of luciferase activity of Panc-1 orthotopic tumors

at day 35 and day 84, post tumor induction. Control Exo (n=5), MSCs siKrasG12D-1 iExo,

CB (n=5), MSCs siKrasG12D-1 iExo, RB (n=4), BJ siKrasG12D-1 iExo, RB (n=5). (D) Panc-1

orthotopic tumor growth (bioluminescence) over time. Control Exo (n=5), MSCs

siKrasG12D-1 iExo, CB (n=5), MSCs siKrasG12D-1 iExo, RB (n=4), BJ siKrasG12D-1 iExo, RB

(n=5). (E) Tumor burden (%) of Panc-1 tumor bearing mice at experimental endpoint.

Control Exo (n=5), MSCs siKrasG12D-1 iExo, CB (n=5), MSCs siKrasG12D-1 iExo, RB (n=4),

BJ siKrasG12D-1 iExo, RB (n=5). (F) Tumor weights (g: grams) of Panc-1 tumor bearing

mice at experimental endpoint. Control Exo (n=5), MSCs siKrasG12D-1 iExo, CB (n=5),

MSCs siKrasG12D-1 iExo, RB (n=4), BJ siKrasG12D-1 iExo, RB (n=5). (G) Flow cytometry

analyses and quantification of apoptosis in Panc-1 cells comparing the efficacy of 48

distinct electroporation conditions (A1-A16; B1-B16; C1-C16) of MSCs and BJ Fibroblast

siKrasG12D iExosomes, using a large volume electroporation system and CB (n=2 distinct

experiments). The electroporation condition subsequently chosen was A-14. The mean

+/- SEM is depicted. See accompanying source data.


(A) Schematic representation of the experiment using KPC689 orthotopic tumor bearing

mice (related to experiment in Figure 5E). (B) Comparison of baseline tumor

bioluminescence (total Flux) pre-treatment, of KPC689 orthotopic experiments 1 and 2,

to show baseline tumor measurements pre-treatment of Experiment-2 was higher than

that of Experiment-1. Experiment-1: Control Exo (n=4), BJ siKrasG12D-1 iExo (n=6) and

MSCs siKrasG12D-1 iExo (n=6). Experiment-2: CB/PBS (n=7), Control Exo (n=7),

Gemcitabine (n=8), MSCs siKrasG12D-2 iExo (n=8), Gemcitabine + MSCs siKrasG12D-2 iExo

(n=8). Two-tailed unpaired t test between experiment 1 and 2. (C) Detailed findings from

individual mice with KPC689 tumors (Experiment-2) treated with CB/PBS (n=7), Control

Exo (n=7), Gemcitabine (n=8), MSCs siKrasG12D-2 iExo (n=8), Gemcitabine + MSCs

siKrasG12D-2 iExo (n=8), showing survival post treatment start (PTS) and post birth (PB),

tumor weight (TW) and body weight (BW), number of lung macrometastases, and

observations at necropsy/euthanasia. (D) Representative images of H&E stained

sections of pancreatic tumors from KPC689 tumor bearing mice. Scale bars 100 μm (top)

and 50 μm (bottom). CB/PBS (n=7), Control Exo (n=7), Gemcitabine (n=8), MSCs

siKrasG12D-2 iExo (n=8), Gemcitabine + MSCs siKrasG12D-2 iExo (n=8). The mean +/- SEM

is depicted. See accompanying source data.


(A) Schematic representation of the experiment using KPC689 orthotopic tumor bearing

mice (related to experiment in Figure 5F). (B) Detailed findings from individual mice with

KPC689 tumors treated with CB/PBS (n=7), Control Exo (n=7), Gemcitabine (n=7), MSCs

siKrasG12D-2 iExo (n=7), Gemcitabine + MSCs siKrasG12D-2 iExo (n=7), showing survival

post treatment start (PTS) and post birth (PB), tumor weight (TW) and body weight (BW),

number of lung macrometastases, and observations at necropsy/euthanasia.


(A) Cytotoxicity tests performed on the blood collected from adult C57Bl/6J mice injected

i.p. every other day for 120 days. PBS (n=3), BJ fibroblasts exosomes (n=4). ALT: alanine

aminotransferase; AST: aspartate aminotransferase; BUN: blood urea nitrogen. The grey

highlight defines the normal range. (B) Representative pictures of H&E stained tissue

sections showing lesions of inflammation observed in the mesenteric lymph node, kidney,

liver and gall-bladder.


(A) Immunotyping analyses of spleen, bone marrow and thymus of immunocomptent non-

tumor bearing mice administered with the indicated treatments. ‡: these values are not

erroneous despite their similarities. #: this group shows statistical significant differences

with exosomes treated mice but not with CB. (B) ELISA for IL-6 in the indicated groups.

0 denotes that the levels were below the limits of the assay (not detected).


(A) Biodistribution in the indicated organs of DiR labeled MSCs exosomes 6 hours post

i.v. injection in nude mice (left) or 3 hours post i.v. injection in tumor bearing (KPC689)

C57BL/6 mice. DiR only (n=1), DiR labeled MSCs exosomes (n=2). (B) Biodistribution in

the indicated organs of DiR labeled MSCs exosomes 3 hours post i.p. or i.v. injection of

C57BL/6 non-tumor bearing mice. (C) Biodistribution in the indicated organs of DiR

labeled MSCs exosomes 24 and 48 hours post i.p. injection tumor bearing (KPC689)

C57BL/6 mice. These tumors are depicted in Figure 7B.


(A) Representative dot plot of flow cytometry analyses of apoptosis in Panc-1 cells

induced by MSCs siKrasG12D-2 iExosomes freeze/thaw during 3 and 6 months showing

similar efficiency that fresh iExosomes. n=2 independent experiments, quantitation is

shown in Figure 6I. (B) Detailed findings from individual PKS mice treated with MSCs

siKrasG12D-2 iExo (n=4) and Control Exo (n=5), showing treatment groups, start of

treatment, survival of mice, body weight (BW) and tumor weight (TW) at experimental

endpoint. See accompanying source data.

Supplementary Table 1

Detailed necropsy analyses of C57BL/6 mice injected i.p. with either empty BJ Fibroblast

exosomes (n=4) or PBS (n=3), every other day, for 120 days.

Supplemental Table 2

Summary of treatments for the in vivo experiments.

Supplementary Table 3

Detailed information on the antibodies used in this study.


Panc−

1 iExo−3

Panc−

1 iExo−2

Panc−

1 iExo−1

Panc−

1 con

t−3

Panc−

1 scr−

2

Panc−

1 scr−

1

Panc−

1 con

t−1

Panc−

1 CE−

1

Panc−

1 CE−

3ILMN_1701789ILMN_3242144ILMN_1693483ILMN_2239754ILMN_1791280ILMN_3241369ILMN_3240150ILMN_2165762ILMN_2295879ILMN_1705750ILMN_3240306ILMN_1770994ILMN_1661470ILMN_1716264ILMN_1774973ILMN_1743217ILMN_3244592ILMN_3238478ILMN_1766650ILMN_1662358ILMN_3241140ILMN_1734542ILMN_1658247ILMN_2053527ILMN_1739428ILMN_1795963ILMN_1665554ILMN_1874689ILMN_1724533ILMN_1814305ILMN_1891455ILMN_1682354ILMN_1782167ILMN_2376403ILMN_3248562ILMN_1770030ILMN_1652434ILMN_1754894ILMN_1885233ILMN_1784380ILMN_3247008ILMN_1689294ILMN_1713832ILMN_1730201ILMN_2214355ILMN_3235397ILMN_3241139ILMN_1652638ILMN_1747183ILMN_1660900ILMN_1711120ILMN_1731224ILMN_1698191ILMN_1811866ILMN_1682636ILMN_1741632ILMN_1704056ILMN_1789990ILMN_2370624ILMN_3274045ILMN_1699772ILMN_3241798ILMN_1666757ILMN_1762275ILMN_1728106ILMN_3243644ILMN_1760102ILMN_1795317ILMN_1795338ILMN_1740555ILMN_1794560ILMN_2112730ILMN_1762606ILMN_1667594ILMN_1654429ILMN_1711838ILMN_1739103ILMN_1701487ILMN_3233229ILMN_2181892ILMN_1665832ILMN_1659953ILMN_1810127ILMN_1693401ILMN_2401701ILMN_1711005ILMN_1694367ILMN_1669700ILMN_2209515ILMN_2377829ILMN_1766054ILMN_1911605ILMN_1657348ILMN_2052373ILMN_1745807ILMN_1696846ILMN_1672295ILMN_1675992ILMN_1693242ILMN_1785037ILMN_1787539ILMN_3249167ILMN_1745397ILMN_1810712ILMN_1713351ILMN_1709604ILMN_2376205ILMN_1670970ILMN_1802096ILMN_3220934ILMN_1797596ILMN_1654072ILMN_1701918ILMN_1813685ILMN_1762003ILMN_1867149ILMN_1745148ILMN_1659544ILMN_2123431ILMN_1781373ILMN_1780711ILMN_1685703ILMN_2401618ILMN_1764383ILMN_1806845ILMN_1716265ILMN_1764052ILMN_3237291ILMN_1690125ILMN_1778625ILMN_1741727ILMN_1659463ILMN_1678919ILMN_1683415ILMN_1788832ILMN_3240586ILMN_3251232ILMN_2393046ILMN_1744006ILMN_1655748ILMN_1728083ILMN_1859908ILMN_1690464ILMN_3307892ILMN_1699525ILMN_2381697ILMN_2089167ILMN_3237977ILMN_1771697ILMN_1809483ILMN_1696956ILMN_1772292ILMN_1863592ILMN_1681644ILMN_1654685ILMN_1879482ILMN_1695606ILMN_1667239ILMN_1764851ILMN_2214678ILMN_1764723ILMN_1854833ILMN_1758337ILMN_1722292ILMN_1722781ILMN_1783185ILMN_2388800ILMN_1765274ILMN_2160160ILMN_1738725ILMN_3204890ILMN_1756417ILMN_3234762ILMN_1710544ILMN_1695246ILMN_3200140ILMN_2198878ILMN_1777483ILMN_1712298ILMN_3244157ILMN_1766219ILMN_1703913ILMN_1764166ILMN_1754332ILMN_1815705ILMN_1696622ILMN_1748124ILMN_1657234ILMN_1745154ILMN_2400219ILMN_1803611ILMN_2415144ILMN_2067852ILMN_1716441ILMN_3235185ILMN_2413158ILMN_1795778ILMN_2318685ILMN_2283325ILMN_1680465ILMN_1713846ILMN_1709287ILMN_1652762ILMN_3249435ILMN_2082762ILMN_1714765ILMN_1763000ILMN_1800733ILMN_3242448ILMN_1671837ILMN_1837935ILMN_1708110ILMN_1727855ILMN_2170625ILMN_1756784ILMN_3310840ILMN_1701111ILMN_2226628ILMN_2303955ILMN_1760556ILMN_1798181ILMN_3248781ILMN_1688103ILMN_1856480ILMN_1707051ILMN_1659976ILMN_1712545ILMN_1814151ILMN_1758164ILMN_1792110ILMN_3238283ILMN_3251634ILMN_2348268ILMN_1661363ILMN_1723706ILMN_1693259ILMN_1677376ILMN_1855278ILMN_1754860ILMN_1656129ILMN_1743402ILMN_3265237ILMN_1690442ILMN_1674399ILMN_1663119ILMN_1752478ILMN_1713751ILMN_2149164ILMN_3188106ILMN_1802404ILMN_1705241ILMN_1774265ILMN_1804929ILMN_1706118ILMN_1745420ILMN_1705032ILMN_1745374ILMN_1715814ILMN_2229877ILMN_1733615ILMN_1800942ILMN_1800512ILMN_1669210ILMN_1708778ILMN_1707695ILMN_2063584ILMN_1763745ILMN_2386205ILMN_2385278ILMN_1681984ILMN_1779376ILMN_3182171ILMN_1746673ILMN_2220184ILMN_1745116ILMN_2130514ILMN_2322375ILMN_1662243ILMN_1765825ILMN_1677691ILMN_1779356ILMN_1746819ILMN_1682658ILMN_1666306ILMN_1667825ILMN_2054297ILMN_1747673ILMN_1807873ILMN_1691790ILMN_1710000ILMN_2124155ILMN_3251691ILMN_1718309ILMN_2339705ILMN_2218758ILMN_1704529ILMN_2327795ILMN_1815707ILMN_1678808ILMN_1660079ILMN_1680692ILMN_1784749ILMN_1702691ILMN_2092756ILMN_1810797ILMN_1673548ILMN_1745079ILMN_1729748ILMN_2381296ILMN_1903159ILMN_1701724ILMN_3294134ILMN_1703891ILMN_3251550ILMN_1696946ILMN_3247261ILMN_1706873ILMN_1695590ILMN_2086077ILMN_2148012ILMN_1700231ILMN_2173835ILMN_2347798ILMN_1717639ILMN_2377980ILMN_1667748ILMN_1804051ILMN_1678669ILMN_1765966ILMN_2220403ILMN_1684197ILMN_1814966ILMN_2353642ILMN_1774432ILMN_1763144ILMN_1721833ILMN_1807136ILMN_1785284ILMN_1738712ILMN_1680347ILMN_2179726ILMN_1680364ILMN_1679133ILMN_1787509ILMN_1772959ILMN_2135175ILMN_1746801ILMN_1719975ILMN_1792409ILMN_1779043ILMN_1792910ILMN_3308753ILMN_1724789ILMN_3187328ILMN_1667711ILMN_1667201ILMN_3282587ILMN_2389376ILMN_3298694ILMN_1680091ILMN_1710284ILMN_2348975ILMN_2213199ILMN_1680139ILMN_1697153ILMN_1793549ILMN_1701386ILMN_1713901ILMN_1747556ILMN_1677466ILMN_1729749ILMN_1656574ILMN_2268068ILMN_2327346ILMN_1745034ILMN_1660871ILMN_1779751ILMN_1682812ILMN_1710075ILMN_2183409ILMN_1659937ILMN_1806486ILMN_1779558ILMN_2204754ILMN_1660718ILMN_1680693ILMN_1726104ILMN_1809842ILMN_1659027ILMN_1760121ILMN_1728071ILMN_1717313ILMN_1812926ILMN_3247018ILMN_2134538ILMN_3306173ILMN_2392286ILMN_2198515ILMN_1687867ILMN_1730809ILMN_2325347ILMN_1704385ILMN_1815733ILMN_1788213ILMN_1736847ILMN_1812441ILMN_1773742ILMN_1810069ILMN_1724406ILMN_2390310ILMN_2374115ILMN_1781155ILMN_3190972ILMN_1795826ILMN_1786347ILMN_2246510ILMN_1662065ILMN_3229552ILMN_3310416ILMN_1783156ILMN_1734814ILMN_1674038ILMN_1892638ILMN_1804988ILMN_1756676ILMN_1756999ILMN_3241091ILMN_1673275ILMN_1719286ILMN_1692191ILMN_2120022ILMN_1737949ILMN_1813400ILMN_1665291ILMN_1740604ILMN_1778173ILMN_1790537ILMN_2209578ILMN_1745282ILMN_1774800ILMN_1789384ILMN_1703430ILMN_1667839ILMN_1669584ILMN_1677092ILMN_1696254ILMN_2374159ILMN_1798212ILMN_1702065ILMN_1715324ILMN_1666776ILMN_2367883ILMN_1731720ILMN_2383383ILMN_1660111ILMN_1776047ILMN_2355033ILMN_3233179ILMN_1721704ILMN_1788547ILMN_2119555ILMN_3268697ILMN_1694686ILMN_2075927ILMN_1674302ILMN_2328666ILMN_1803317ILMN_3227023ILMN_1726786ILMN_3244640ILMN_2166524ILMN_1736042ILMN_2231021ILMN_3211887ILMN_1773865ILMN_1673936ILMN_1695475ILMN_1707748ILMN_1683096ILMN_1786041ILMN_2374778ILMN_1700202ILMN_1763228ILMN_1749011ILMN_1723768ILMN_1812297ILMN_2381064ILMN_1794539ILMN_1701131ILMN_3235326ILMN_1693341ILMN_1802205ILMN_1676191ILMN_1685781ILMN_1781636ILMN_1671005ILMN_1799744ILMN_1737604ILMN_1786976ILMN_1720270ILMN_1802615ILMN_2285802ILMN_1806907ILMN_1732071ILMN_2358540ILMN_1664216ILMN_1700584ILMN_1807596ILMN_1663042ILMN_1701457ILMN_1743836ILMN_1794875ILMN_1728228ILMN_1775822ILMN_1735930ILMN_1665865ILMN_1785336ILMN_1791569ILMN_2143155ILMN_1696643ILMN_1746883ILMN_1748827ILMN_1802380ILMN_1705814ILMN_1719870ILMN_1718898ILMN_1734833ILMN_1678904ILMN_1669252ILMN_1717934ILMN_1656196ILMN_1696432ILMN_1705093ILMN_1790534ILMN_1669905ILMN_2220187ILMN_1802355ILMN_1756806ILMN_1661595ILMN_2360730ILMN_1798886ILMN_2374164ILMN_1663866ILMN_1726288ILMN_1664303ILMN_1720858ILMN_1771333ILMN_3308961ILMN_1771841ILMN_1664068ILMN_2148847ILMN_1655469ILMN_1719695ILMN_1792681ILMN_1745471ILMN_1667213ILMN_1794501ILMN_1794726ILMN_1732688ILMN_1780582ILMN_1681737ILMN_1713764ILMN_2154322ILMN_2385220ILMN_1717056ILMN_1693620ILMN_1700690ILMN_2405018ILMN_1690105ILMN_1780302ILMN_1659122ILMN_1770811ILMN_1689378ILMN_1763837ILMN_1753183ILMN_1756992ILMN_2157510ILMN_1898723ILMN_1682799ILMN_1684289ILMN_1664466ILMN_2147114ILMN_1787815ILMN_1737728ILMN_2059294ILMN_2386100ILMN_1761247ILMN_1729033ILMN_1677814ILMN_1726460ILMN_2376408ILMN_1691111ILMN_2214997ILMN_1660554ILMN_1657153ILMN_1671603ILMN_1728972ILMN_1792455ILMN_1807719ILMN_1787844ILMN_1668012ILMN_1698209ILMN_1790549ILMN_1683891ILMN_1666733ILMN_2411897ILMN_1664912ILMN_1685916ILMN_1748018ILMN_1682775ILMN_1655990ILMN_1671557ILMN_1777794ILMN_1808354ILMN_3223500ILMN_1661599ILMN_1792138ILMN_1793616ILMN_1679093ILMN_1683859ILMN_1670752ILMN_1787897ILMN_1699354ILMN_3237404ILMN_1787691ILMN_1764500ILMN_2086105ILMN_3231881ILMN_3214893ILMN_2146372ILMN_2150465ILMN_1687375ILMN_2122511ILMN_2186061ILMN_2155228ILMN_1687583ILMN_1793672ILMN_1758658ILMN_1712112ILMN_3270641ILMN_1698148ILMN_1838313ILMN_1729234ILMN_1724145ILMN_1675448ILMN_1700276ILMN_1675239ILMN_3227315ILMN_1745806ILMN_1711702ILMN_2320250ILMN_3237177ILMN_1715401ILMN_2415979ILMN_1667430ILMN_1720158ILMN_1815319ILMN_1679405ILMN_1735453ILMN_1732452ILMN_1757497ILMN_1651705ILMN_1661593ILMN_1804610ILMN_1669046ILMN_1665455ILMN_1668507ILMN_1717674ILMN_2045994ILMN_1701998ILMN_2084353ILMN_1805519ILMN_2173294ILMN_2338963ILMN_2397954ILMN_2350574ILMN_3251737ILMN_1791576ILMN_3238680ILMN_3274596ILMN_2404135ILMN_3239861ILMN_2222008ILMN_1803988ILMN_1755909ILMN_1793476ILMN_3248773ILMN_1808938ILMN_1727194ILMN_1674551ILMN_3197767ILMN_1815308ILMN_1758811ILMN_2054019ILMN_1759954ILMN_1768973ILMN_3181480ILMN_1763688ILMN_2398107ILMN_3288717ILMN_2181432ILMN_1751072ILMN_2219618ILMN_1777740ILMN_3279414ILMN_1719039ILMN_1703697ILMN_1746525ILMN_1652379ILMN_2144426ILMN_3272603ILMN_2091347ILMN_3305304ILMN_1726783ILMN_1737314ILMN_1682738ILMN_1768101ILMN_1814122ILMN_2231020ILMN_1898124ILMN_1664516ILMN_1681016ILMN_1672504ILMN_1706502ILMN_3229324ILMN_2110908ILMN_1671554ILMN_2180371ILMN_1776109ILMN_1796074ILMN_1778836ILMN_1666453ILMN_1796177ILMN_1654920ILMN_2380588ILMN_1775304ILMN_1780236ILMN_1708006ILMN_2096784ILMN_1770053ILMN_2224907ILMN_1668721ILMN_3206804ILMN_2365465ILMN_3305938ILMN_1658847ILMN_1677607ILMN_1716446ILMN_1662340ILMN_2353358ILMN_1695706ILMN_1756220ILMN_2364521ILMN_2057573ILMN_2367239ILMN_1664756ILMN_1777325ILMN_2294762ILMN_1740938ILMN_1679796ILMN_1794017ILMN_1723971ILMN_1693270ILMN_2374692ILMN_1652128ILMN_1752631ILMN_2342437ILMN_1703946ILMN_2192694ILMN_1790778ILMN_1669523ILMN_2061565ILMN_2348788ILMN_1811104ILMN_2136089ILMN_1672004ILMN_1791896ILMN_1746408ILMN_2089329ILMN_1707503ILMN_1653856ILMN_1755405ILMN_1674661ILMN_1755354ILMN_1814971ILMN_1797964ILMN_1669113ILMN_1708375ILMN_1791296ILMN_2215382ILMN_1744381ILMN_1805192ILMN_1698213ILMN_1669456ILMN_1699856ILMN_1812769ILMN_1706013ILMN_1733519ILMN_3242900ILMN_1695827ILMN_2313889ILMN_2339955ILMN_1803824ILMN_3289352ILMN_1720282ILMN_1772821ILMN_3310351ILMN_1654268ILMN_1750278ILMN_1778611ILMN_1660793ILMN_1673113ILMN_1738150ILMN_1703123ILMN_3239181ILMN_1696911ILMN_2061435ILMN_1700681ILMN_2356654ILMN_1747016ILMN_1718063ILMN_1732296ILMN_1659047ILMN_2285713ILMN_1784709ILMN_1759915ILMN_1776105ILMN_2092536ILMN_2405684ILMN_1673305ILMN_3268165ILMN_1809433ILMN_2113738ILMN_1763091ILMN_1719749ILMN_1780058ILMN_1737588ILMN_1706015ILMN_1691364ILMN_2230902ILMN_1740426ILMN_1727479ILMN_2125675ILMN_2349459ILMN_1687384ILMN_1664861ILMN_2099301ILMN_1674620ILMN_1658289ILMN_1801939ILMN_1702487ILMN_2070052ILMN_2049727ILMN_1791226ILMN_2094776ILMN_3239135ILMN_2200917ILMN_1760667ILMN_2286014ILMN_1702168ILMN_1692145ILMN_1778561ILMN_1797776ILMN_1776080ILMN_1671893ILMN_2063168ILMN_2136133ILMN_1782305ILMN_1659936ILMN_1805737ILMN_2395043ILMN_2041190ILMN_1803811ILMN_1695420ILMN_1685480ILMN_2073235ILMN_1720889ILMN_3214256ILMN_2404850ILMN_2285996ILMN_3205271ILMN_1701877ILMN_3304111ILMN_1776181ILMN_1665823ILMN_2396020ILMN_1737406ILMN_2184373ILMN_2347068ILMN_1776347ILMN_1763540ILMN_1658835ILMN_1749662ILMN_1680618ILMN_1707727ILMN_3305169ILMN_1806692ILMN_2194852ILMN_1655710ILMN_2051900ILMN_1656501ILMN_3206343ILMN_1775170ILMN_3187612ILMN_3244117ILMN_2188119ILMN_1795930ILMN_1683664ILMN_1684306ILMN_1786242ILMN_1761804ILMN_3202024ILMN_1784602ILMN_1795937ILMN_1687887ILMN_3251472ILMN_2134381ILMN_2408415ILMN_2345837ILMN_1742163ILMN_3237656ILMN_1776052ILMN_2091123ILMN_1803398ILMN_1688780ILMN_1804396ILMN_1748591ILMN_1706687ILMN_2041046ILMN_2192693ILMN_3225591ILMN_1796417ILMN_1736597ILMN_3278506ILMN_1811468ILMN_1737184ILMN_1757467ILMN_1673711ILMN_1769705ILMN_3178792ILMN_1713892ILMN_1656057ILMN_3264112ILMN_1668052ILMN_1688480ILMN_1765446ILMN_1668411ILMN_1773337ILMN_1806023ILMN_1711862ILMN_2313730ILMN_2115125ILMN_3275345ILMN_2094106ILMN_1724040ILMN_1731619ILMN_2130078ILMN_2167922ILMN_1711823ILMN_1743199ILMN_1756326ILMN_1805028ILMN_3208715ILMN_1781285ILMN_1762899ILMN_1660577ILMN_2058141ILMN_1716678ILMN_1735014ILMN_1746252ILMN_1689327ILMN_1869109ILMN_3299478ILMN_1697469ILMN_2156267ILMN_1682717ILMN_1751607ILMN_2149226ILMN_1687978ILMN_1675848ILMN_2234016ILMN_1665510ILMN_2308849ILMN_3300797ILMN_1713636ILMN_3302177ILMN_1768534ILMN_2060413ILMN_1773154ILMN_1803429ILMN_1814823ILMN_3176403ILMN_2188264ILMN_1699829ILMN_1740429ILMN_1686664ILMN_1691156ILMN_1753342ILMN_1720829ILMN_1770338

−2 0 1 2Row Z−Score

Color Key

BXPC−3 s

cr−1

BXPC−3 s

cr−3

BXPC−3 s

cr−2

BXPC−3 c

ont−3

BXPC−3 c

ont−1

BXPC−3 c

ont−2

BXPC−3 i

EXO−3

BXPC−3 C

E−1

BXPC−3 C

E−2

BXPC−3 C

E−3

BXPC−3 i

EXO−2

BXPC−3 i

EXO−1

−2 0 2Row Z−Score

Color Key

BXPC

−3 C

E−1

BXPC

−3 s

cr−3

BXPC

−3 s

cr−2

BXPC

−3 s

cr−1

BXPC

−3 c

ont−

3

BXPC

−3 c

ont−

2

BXPC

−3 c

ont−

1

BXPC

−3 C

E−2

BXPC

−3 C

E−3

BXPC

−3 iE

XO−2

BXPC

−3 iE

XO−1

BXPC

−3 iE

XO−3


Color Key

Panc−1

iExo−3

Panc−1

iExo−2

Panc−1

iExo−1

Panc−1

con

t−3

Panc−1

scr−2

Panc−1

scr−1

Panc−1

con

t−1

Panc−1

CE−

1

Panc−1

CE−

3


Color Key

A D

C E

F

NES=2.038FDR=0.0053

NES=2.0382FDR=0.0058

NES=2.034FDR=0.0065

NES=-2.0005FDR=0.0096 NES=-2.0629

FDR=0.0026

NES=-2.2036FDR≃0.0

NES=-2.234FDR≃0.0

NES=-2.0402FDR=0.0039

BxPC-3

−0.2 −0.1 0.0 0.1 0.2 0.3

−0.1

5−0

.10

−0.0

50.

000.

050.

10

Leading logFC dim 1

Lead

ing

logF

C d

im 2

BXPC3_Scr

BXPC3_Scr

BXPC3_Scr

BXPC3_CtrlExoBXPC3_CtrlExo

BXPC3_CtrlExo

BXPC3_sirnaExo

BXPC3_sirnaExo

BXPC3_sirnaExo

BXPC3_Ctrl

BXPC3_CtrlBXPC3_Ctrl

Panc-1

−0.4 −0.2 0.0 0.2 0.4 0.6 0.8

−0.1

0.0

0.1

0.2

0.3

Leading logFC dim 1

Lead

ing

logF

C d

im 2

Panc1_Scr

Panc1_Scr

Panc1_CtrlExo

Panc1_CtrlExoPanc1_sirnaExo

Panc1_sirnaExo

Panc1_sirnaExo

Panc1_Ctrl

Panc1_Ctrl

NES=2.1315FDR=0.0019

NES=2.1518FDR=0.0022

NES=2.1214FDR=0.0017

B


Bowel Kidney Lung

Spleen Liver Pancreas

G G T

G A/G T

G A/G T

Tumor A2-rsi

Tumor B1-0scr0

200

400

600

Tum

or v

olum

e (m

m3 )

siScrbl siKrasG12D-1

US

MRI

A BM

ouse

1M

ouse

2

C

D

Survival (Days) Pre-treatment Post-treatment Post-treatment Post-treatment End-point Observations Notes

Mouse ID Treatment PTS PB Tumor volume (mm3) (US)

Tumor volume (mm3) (MRI)

BW(g) MRI-2 MRI-3 MRI-4 TW(g) BW(g) Necropsy notesSuspected

tumor burden related death

A1-Y siKrasG12D iExo 133 286 211.87 239.56 23.4 CNBM CNBM Euthanized 0.87 28.8 Jaundice, very small tumor burden No

A1-B siKrasG12D iExo 316 469 94.49 112.27 24.6 415.44 146.24 26.98 0.38 31.9 Hunched, swollen intestine, kidney atrophy, mesenteric abscess, minimal tumor burden

No

A1-G siKrasG12D iExo 257 410 86.84 108.52 22.1 515.05 126.97 10.38 0.2 23.8 Hunched, kidney atrophy, minimal tumor buden No

A1-O siKrasG12D iExo 360 513 100.08 167.31 25.3 363.45 150.16 63.84 0.24 24.2Enlarged mesenteric lymph node, minimal tumor

burden No

A1-W siKrasG12D iExo 254 407 225.92 231.50 23.6 CNBM 212.13 50.44 0.28 28.1Enlarged eyes, euthanasia due to neurological

problems, small tumor No

A2-Y siKrasG12D iExo 35 188 434.96 481.62 22.1 642.22 Euthanized Euthanized 1.4 29.2 Moderate tumor burden, euthanized as an age matched comparison to B1-O

N/A: age-matched

A2-NT siKrasG12D iExo 178 331 212.71 206.30 25.1 CNBM 965.71 CNBM 1.8 32.2Small primary tumor, large secondary tumor,

euthanized due to tumor burden Yes

Average: 195.27 221.01 484.04 320.24 37.91 0.74

B1-Y siScrbl iExo 206 359 158.60 186.11 25.0 CNBM 654.78 Euthanized 1.21 26.1Moderate tumor burden, hunched, scaly skin at

euthanasia Yes

B1-B siScrbl iExo 168 321 114.13 112.30 24.4 773.19 805.08 Euthanized 1.61 26.0 Moderate tumor burden, weak, swollen bowel YesB1-G siScrbl iExo 144 297 98.81 146.54 23.1 961.41 1977.47 Euthanized 5.84 27.9 Extensive tumor burden, hunched posture, weak Yes

B1-O siScrbl iExo 35 188 447.43 379.32 22.9 1254.40 Euthanized Euthanized 2.3 31.2Extensive tumor burden, euthanized due to tumor

burden, aged matched with A2-Y Yes

B1-W siScrbl iExo 102 255 176.84 182.64 24.8 CNBM 2225.12 Euthanized 4.67 34.1Large primary and secondary tumors, spleen

invasion, macroscopic liver nodulesN/A: age-matched

B2-Y siScrbl iExo 158 311 250.26 263.35 26.1 950.14 CNBM Euthanized 2.4 28.8 Extensive tumor burden, weak, hunched posture YesB2-NT siScrbl iExo 69 222 86.75 120.45 25.4 CNBM 969.73 Euthanized 1.69 29.2 Weak, moderate tumor burden, spleen atrophy Yes

Average: 190.40 198.68 984.79 1326.43 2.82


100% 100% 96%

100%

100%

0.8% 0.1% 0.2%

47%86%

100%100%

0.2%

84%

3.6%

100% 100% 81%

0.1%

94%

83%

100% 100% 93%

99%

98.1%

99%

3.6% 0.5%0%

B C

CD44 CD90 CD105

CD73 HLA-ABCCD29

CD9 CD63CD81

CD47 CD45CD10

HLA-DR CD34CD31 HLA-DR CD34CD31

CD47 CD45CD10

CD9 CD63CD81

CD73 HLA-ABCCD29

CD44 CD90 CD105

% o

f Max

% o

f Max

A

Donor 1 Donor 2 Donor 30

500

1000

1500

2000

2500

3000N

umbe

r of e

xoso

mes

per

cel

l per

hou

r

*

Day 0 Day 1 Day 2

Day 3

Day 4

Day 5

Day 6

Day 7

Day 8

Day 9

Preparation

of bioreactor

Seeding

of

MSCs

Expansion of MSCs in growth media Wash with PBS and

load PLT-free media

A Timeline for MSCs-derived exosomes production using a bioreactor

Storage of MSCs iExo

aliquots at –80∘C

Thaw/Pool of

conditioned

media

Filtration

(0.2 µm)

Resuspension of MSCs-

derived exosomes in

research buffer (RB) or

clinical buffer (CB)

Ultracentrifugation

100,000g for 3 h

Electroporation

of exosomes

with KrasG12D

siRNA (iExo)

B Isolation of and generation of MSCs-derived iExosomes

C Generation of GMP grade MSCs-derived iExosomes

Day 11 Day 13 Day 15 Day 17 Day 19 Day 21

Bioreactor

Harvest 1

↓

Freeze (-80∘C)

Bioreactor

Harvest 2

↓

Freeze (-80∘C)

Bioreactor

Harvest 3

↓

Freeze (-80∘C)

Bioreactor

Harvest 4

↓

Freeze (-80∘C)

Bioreactor

Harvest 5

↓

Freeze (-80∘C)

Bioreactor

Harvest 6

↓

Freeze (-80∘C)


Characterization of

exosomes by NanoSightTM

(30nm-250nm) and surface

markers (CD9, CD47,

CD29, CD105) + sterility

test

Dose preparations of

siRNA MSCs

exosomes for

infusion into patients

Day 0-19 Day 23-25

Pool of MSC-

conditioned

media and

centrifugation 1,000gfor 15 min at 4∘C

High speed

centrifugation

100,000g

Resuspension

of exosomes

+Exosomes siRNA

Day 40

Exosomes

carrying siRNA

Growth of MSC and

collection of

conditioned media

from the bioreactor

0.2 µm filter

Exosomes

pellet

Exosomes

pellet

(Sterility test from

each harvests)

Bioreactor

(Terumo BCT)

Cobe 2991 Cell

Processor

(Terumo BCT)

Bioreactor harvests:

Harvest time(days)

Volume(ml)

Protein (mg)

Exosomes (x109)

Harvest 1 10 250 2.641 2010

Harvest 2 12 250 6.222 4493

Harvest 3 14 250 3.606 3402

Harvest 4 16 250 3.534 3472

Harvest 5 18 250 0.992 983

Harvest 6 20 250 1.040 1210

C

0 5 10 15 2002468

10

80

100

120

140

Days

Leve

ls o

f glu

cose

and

lact

ose

Glucose (mg/dL)Lactose (mmol/L)

1 2 30

5

10

15

20

Num

ber o

f exo

som

es (1

012 )

Bioreactor run

BA

D

Harvest 1 Harvest 2

Harvest 3 Harvest 4

Harvest 5 Harvest 6

78.1% 85.0%84.6% 99.3%

97.2% 98.3%99.4%

92.9% 97.3%82.0% 80.0%

81.7%

93.2%

CD9 CD63 CD81 CD47

CD9 CD63 CD81 CD47

CD9 CD63 CD81 CD47

99.3%

99.0%

100% 100% 99.9%

100% 100% 99.9%

99.9% 95.3%

CD9 CD63 CD81 CD47

CD9 CD63 CD81 CD47

CD9 CD63 CD81 CD47

% o

f Max

% o

f Max

% o

f Max

97.1%


0 10 20 30 40 50 600

1×107

2×107

3×107

4×107

Days (Post tumor induction)

Tota

l flux

(p/s

ec)

Control ExoBJ siKrasG12D-1 iExoMSCsiKrasG12D-1 iExo

Treatment Start

Supplemental Figure 6P

an

cre

as

Liv

er

Sp

lee

nK

idn

ey

Control Exo MSCs siKrasG12D-1 iExoBJ siKrasG12D-1 iExo

Contro

l Exo

BJ siK

rasG

12D-1 iE

xo

MSCs siK

rasG

12D-1 iE

xo0

5

10

15

Tum

or w

eigh

t at

ex

perim

enta

l en

dpoi

nt (g

)

Contro

l Exo

BJ siK

rasG

12D-1 iE

xo

MSCs siK

rasG

12D-1 iE

xo0

10

20

30

40

Tum

or b

urde

n at

ex

perim

enta

l en

dpoi

nt (%

)

E F

B

C

G

D

0

25

50

75

100

% o

f mic

e

Macrometastases/local invasiion detectedNo macrometastasis/local invasion detected

splee

nlun

gliv

er

Control Exo (n=4)

MSC siKrasG12D-1 iExo (n=6)

diaph

ragmkid

ney

splee

nlun

gliv

er

diaph

ragmkid

ney

splee

nlun

gliv

er

diaph

ragmkid

ney

BJ siKrasG12D-1 iExo (n=6)

Survival (Days) Pre-treatment End-point Observations Notes

Treatment PTS PB BW(g) TW(g) BW(g) Lung macro metastases Necropsy notes Suspected tumor

burden related deathMSCs Ctrl Exo 44 107 20.50 4.81 23.11 59

Large primary and secondary tumors, spleen invasion, liver invasion, diapragm invasion, macroscopic lung nodules

Yes

MSCs Ctrl Exo 37 100 20.00 7.00 23.90 49Large primary and secondary tumors, spleen invasion, diapragm

invasion, kidney invasion, macroscopic lung nodulesYes

MSCs Ctrl Exo 37 100 21.30 5.34 24.12 42Large primary and secondary tumors, diapragm invasion, macroscopic

lung nodulesYes

MSCs Ctrl Exo 44 107 21.60 6.22 27.26 76Large primary and secondary tumors, spleen invasion, liver invasion,

diapragm invasion, macroscopic lung nodulesYes

Average: 5.84BJ siKrasG12D-1 iExo 37 100 20.80 2.14 23.20 35 Moderate tumor, spleen invasion, macroscopic lung nodules Yes

BJ siKrasG12D-1 iExo 49 112 20.20 3.18 25.89 0 Moderate tumor, hunched, weak Yes

BJ siKrasG12D-1 iExo 66 129 20.50 7.92 31.24 0 Moderate tumor burden, hunched, spleen invasion, liver invasion Yes

BJ siKrasG12D-1 iExo 75 138 19.70 2.41 22.87 2Moderate tumor burden, spleen invasion, necrosis of primary tumor,

macroscopic lung nodulesYes

BJ siKrasG12D-1 iExo 101 164 20.30 6.40 28.31 0 Moderate tumor burden, hole in primary tumor hunched, weak Yes

BJ siKrasG12D-1 iExo 120 183 19.80 5.43 32.64 5 Moderate tumor burden, macroscopic lung nodules Yes

Average: 4.58MSCs siKrasG12D-1 iExo 54 117 21.30 1.62 26.40 0 Small tumor, no invasion No

MSCs siKrasG12D-1 iExo 54 117 21.60 2.76 31.86 0 Small tumor, no invasion, some internal bleeding Yes

MSCs siKrasG12D-1 iExo 75 138 19.70 5.64 23.00 0 Moderate tumor burden, no invasion, necrosis of primary tumor Yes

MSCs siKrasG12D-1 iExo 78 141 19.80 2.46 22.93 0 Moderate tumor, tumour in thoracic cavity, necrosis of primary tumor Yes

MSCs siKrasG12D-1 iExo 82 145 21.00 4.01 33.46 6Moderate tumor, necrosis of primary and secondary (subcutaneal) tumors, spleen invasion, kidney invasion, macroscopic lung nodules

Yes

MSCs siKrasG12D-1 iExo 115 178 21.00 11.00 35.48 32Moderate tumor burden, spleen invasion, kidney invasion, diagraphm

invasion, macroscopic lung nodulesYes

Average: 4.58

A

Untrea

ted

Contro

l Exo

MSC siKra

sG12

D-1 iExo

0

10

20

30

40

50P

erce

ntag

e of

apo

ptot

ic c

ells **

Figure 6BUntreated LS HS

13.2 78.0 86.312.9 90.0 89.011.1 84.0 82.0

Figure 6C

Untreated 11.19 8.69 9.48 1.00 1.00 1.00LS 13.28 11.10 12.76 0.23 0.19 0.10HS 13.20 12.95 13.50 0.25 0.05 0.06

Figure 6DsiKrasG12D-1 input siKrasG12D-2 input MSC siKrasG12D-1 iExo, HS, CB MSC siKrasG12D-1 iExo, HS, CB + RN MSC siKrasG12D-1 iExo, HS, CB + T + RN MSC siKrasG12D-1 iExo, LS, RB BJ siKrasG12D-1 iExo, LS, RB MSC siKrasG12D-2 iExo, LS, RB

0.05304851 0.05296884 0.05365822 0.05240104 0.04080845 0.05203353 0.05106859 0.051063970.05272926 0.05380001 0.03918899 0.05270242 0.05184339 0.051577130.05333581 0.05158886 0.04099962 0.05054021 0.05079002 0.05264135

Figure 6E, F: see Supplemental Figures 8C and 9B, respectively.

Delta Ct fold

Control Exo

BJ siKrasG12D iExo

Research Buffer (RB)

MSC siKrasG12D iExo

Research Buffer (RB)

MSCs siKrasG12D iExo

Clinical Buffer (CB)


EB

F

Day 3

5D

ay 8

4

Control

Exo

BJ iExo

RB

MSCs iExo

RB

MSCs iExo

CB

0 20 40 60 80 100 120 140 160 1800.0

5.0×105

1.0×106

1.5×106

2.0×106

2.5×106

3.0×106

Days (Post tumor induction)

Rad

ianc

e (p

/sec

/cm

2 /sr

)

Control ExoBJ iExo RBMSCs IExo RBMSCs iExo CB

Start Treatment

Stop Treatment

C D

A

3 out 5

mice alive

4 out 4

mice alive5 out 5

mice alive

Contro

l Exo

BJ iExo

RB

MSCs iExo

RB

MSCs iExo

CB

0

2

4

6

8

Tum

or w

eigh

t at

expe

rimen

tal

endp

oint

(g)

Contro

l Exo

BJ iExo

RB

MSCs iExo

RB

MSCs iExo

CB

0

5

10

15

20

Tum

or b

urde

n at

ex

perim

enta

l en

dpoi

nt (%

)B

J iE

xo

MS

Cs

iExo

C

ontro

l Exo

Unt

reat

ed B1

B2

B3

B4

B5

B6

B7

B8

B9

B10

B11

B12

B13

B14

B15

B16

0

20

40

60

80

100

Per

cent

age

of a

popt

otic

cel

ls

BJ

iExo

M

SC

s iE

xo

Con

trol E

xoU

ntre

ated C

1C

2C

3C

4C

5C

6C

7C

8C

9C

10C

11C

12C

13C

14C

15C

16

0

20

40

60

80

100

Per

cent

age

of a

popt

otic

cel

ls

BJ

iExo

M

SC

s iE

xo

Con

trol

Exo

Unt

reat

ed A1

A2

A3

A4

A5

A6

A7

A8

A9

A10

A11

A12

A13

A14

A15

A16

0

20

40

60

80

100

Per

cent

age

of a

popt

otic

cel

lsG

Survival (Days) Pre-treatment End-point ObservationsTreatment PTS PB BW(g) TW(g) BW(g) Necropsy notes

MSCs Ctrl Exo 70 133 29.40 1.89 23.00 Small tumor, hunched, diarrhea, no invasion

MSCs Ctrl Exo 71 134 31.10 2.96 33.26 Moderate tumor burden, spleen and liver invasion, ascistes

MSCs Ctrl Exo 85 148 28.70 2.03 30.57 Moderate tumor burden, diaphragm, spleen and kidney invasion

MSCs Ctrl Exo 99 162 34.90 3.26 36.22Moderate tumor burden, abdominal inflammation, ascistes, spleen

and diaphragm invasion

MSCs Ctrl Exo 113 176 25.50 3.10 28.97 Moderate tumor bourden, diaphragm invasion, ascistis

Average: 2.65BJ siKrasG12D-1 iExo RB 113 176 29.30 0.30 25.00 No tumor, hunched posture, diarrhea, no invasion

BJ siKrasG12D-1 iExo RB 143 206 28.20 2.83 27.21 Moderate tumor burden, hunched posture weak, no invasion

BJ siKrasG12D-1 iExo RB 145 208 29.00 4.05 38.50 Moderate tumor burden, hepatomegaly and splenomegaly

BJ siKrasG12D-1 iExo RB 165 228 27.80 2.36 28.00 Moderate tumor burden, spleen invasion

BJ siKrasG12D-1 iExo RB 221 284 31.80 2.54 28.20 Moderate tumor burden,diaprahm, liver and spleen invasion

Average: 2.42

MSCs siKrasG12D-1 iExo RB 165 228 31.30 1.47 21.00Severe abdominal inflammation, diapragm and kidney invasion,

mesenteric metastases

MSCs siKrasG12D-1 iExo RB 145 208 33.00 6.27 36.00 Large primary tumor, diaphragm, liver and spleen invasion

MSCs siKrasG12D-1 iExo RB 130 193 32.00 3.39 28.25 Moderate tumor burden, no invasion

MSCs siKrasG12D-1 iExo RB 198 261 27.90 0.5 25.00 Small tumor burden, no invasion

Average: 2.91BJ siKrasG12D-1 iExo CB 111 174 31.50 1.15 21.30 Moderate tumor burden, diaphragm invasion

BJ siKrasG12D-1 iExo CB 117 180 27.40 1.24 34.40 Moderate tumor, no invasion, ascistes

BJ siKrasG12D-1 iExo CB 221 284 30.40 0.31 18.00No tumor, hunched posture, weak, diarrhea, severe weight loss, no

invasion

BJ siKrasG12D-1 iExo CB 226 289 28.60 2.29 32.71 Moderate tumor burden, diaphragm and, liver invasion

BJ siKrasG12D-1 iExo CB 335 398 27.50 2.27** 30.59

Mouse did not reach moribundancy but showed decreased activity.

The pancreas appears normal yet a mesentaric tumor is noted

together with invasion in the kidneys.

Average: 1.25

Contro

l Exo

BJ siK

rasG

12D-1 iE

xo

MSC siKra

sG12

D-1 iExo

Contro

l Exo

PBS/Plas

malyte

Gemcit

abine

MSC siKra

sG12

D-2 iExo

Gemcit

abine

+MSC si

KrasG

12D-2 iE

xo0

2×106

4×106

6×106

8×106

1×107

Tota

l flux

(p/s

ec)

KPC689Experiment-1

KPC689Experiment-2

* Supplemental Figure 8

PBS/CB buffer MSC siKrasG12D-2 iExo Control Exo GemcitabineGemcitabine +

MSC siKrasG12D-2 iExo D

B

C

0 29 50 76

Time (days)

Tumor implantation

Treatment windows

Gem or PBS

Control Exo or iExo

A

Survival (Days) Pre-treatment End-point Observations

Treatment PTS PB BW(g) TW(g) BW(g) Lung Metastases Necropsy notes

CB/PBS 32 95 21.20 3.26 24.00 0 Moderate tumor burden, diaphragm invasionCB/PBS 34 97 20.50 4.90 21.97 4 Moderate tumor burden, spleen and diaphragm invasion, macroscopic lung nodules

CB/PBS 35 98 19.30 2.55 18.22 0 Moderate tumor burden, peritoneal, diaphragm, spleen iand kidney invasion, macroscopic lung nodules

CB/PBS 35 98 20.90 2.86 20.40 0 Moderate tumor burden, diaphragm invasionCB/PBS 41 104 20.78 2.14 20.78 6 Moderate tumor burden, peritoneal and diaphragm invasion, macroscopic lung nodulesCB/PBS 39 102 19.70 3.75 18.85 0 Moderate tumor burden, spleen, liver and kidney invasionCB/PBS 41 104 21.50 3.26 24.02 0 Moderate tumor burden, diaphragm invasionAverage: 3.25

MSCs Ctrl Exo 42 105 21.10 3.72 20.20 0 Moderate tumor burden, spleen, diaprahm invasion

MSCs Ctrl Exo 39 102 19.80 5.10 23.60 8 Moderate tumor burden, spleen, kidney, liver and diaprahm invasion, macroscopic lung nodules

MSCs Ctrl Exo 50 113 18.50 6.32 24.98 >200 Moderate tumor burden, spleen, kidney and diaprahm invasion, macroscopic lung nodules

MSCs Ctrl Exo 46 109 19.30 3.67 19.70 2 Moderate tumor burden, spleen and kidney invasion, macroscopic lung nodulesMSCs Ctrl Exo 34 97 20.90 2.86 17.50 0 Moderate tumor burden, spleen, liver and diaphragm invasionMSCs Ctrl Exo 36 99 21.30 2.83 22.80 0 Moderate tumor burden, spleen and diaphragm invasionMSCs Ctrl Exo 32 95 18.40 4.52 19.70 2 Moderate tumor burden, diaphragm invasion, macroscopic lung nodulesAverage: 4.15

Gemcitabine 39 102 18.70 0.54 23.47 0 Very small tumor, diaphragm invasion

Gemcitabine 41 104 20.20 2.54 24.66 1 Moderate tumor burden, spleen, kidney and diaphragm invasion, macroscopic lung nodules

Gemcitabine 50 113 21.90 2.3 18.00 0 Moderate tumor burden, no invasionGemcitabine 44 107 22.20 4.77 28.05 0 Moderate tumor burden, spleen, kidney and diaphragm invasion, ascitesGemcitabine 60 123 20.20 0.57 13.25 0 Small tumor, diaphragm invasionGemcitabine 46 109 19.10 1.5 27.20 0 Moderate tumor burden, no invasionGemcitabine 60 123 20.90 2.73 20.07 0 Moderate tumor burden, no invasionGemcitabine 57 120 20.40 3.1 22.90 0 Moderate tumor burden, no invasionAverage: 2.26

MSCs siKrasG12D-2 iExo CB 75 138 19.70 5.83 24.26 11 Moderate tumor burden, liver invasion, macroscopic lung nodulesMSCs siKrasG12D-2 iExo CB 45 108 22.40 1.96 20.00 13 Moderate tumor burden, spleen and diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB 45 108 18.80 1.78 16.15 29 Moderate tumor burden, spleen, kidney and diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB 44 107 20.60 1.72 18.87 0 Moderate tumor burden, spleen, kidney, peritoneal and diaphragm invasion

MSCs siKrasG12D-2 iExo CB 76 139 21.70 5.3 23.67 48 Moderate tumor burden, spleen, kidney and small intestine invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB 44 107 21.30 1.98 24.97 0 Moderate tumor burden, spleen, kidney, liver and diaphragm invasionMSCs siKrasG12D-2 iExo CB 36 99 21.00 1.34 21.60 0 Moderate tumor burden, diaphragm invasionMSCs siKrasG12D-2 iExo CB 44 107 18.90 1.86 20.00 15 Small tumor, diaphragm invasion, lymph node metastases, macroscopic lung nodules

Average: 2.72MSCs siKrasG12D-2 iExo CB+Gem 67 130 20.20 3.32 23.84 >200 Moderate tumor burden, spleen and kidney invasion, macroscopic lung nodulesMSCs siKrasG12D-2 iExo CB+Gem 46 109 20.60 1.14 19.85 0 Small tumor, hunched posture, liver invasion

MSCs siKrasG12D-2 iExo CB+Gem 75 138 19.20 2.27 21.00 8 Moderate tumor burden, spleen, kidney, liver and diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB+Gem 70 133 19.20 3.87 23.84 11 Moderate tumor burden, spleen, kidney and diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB+Gem 50 113 21.00 4.56 23.00 71 Moderate tumor burden, ascites, spleen, kidney and diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB+Gem 50 113 18.40 1.73 19.84 0 No tumor, hunched posture, weak, diarrhea, weight loss, no invasion

MSCs siKrasG12D-2 iExo CB+Gem 95 158 20.20 3.10 25.00 10 Moderate tumor burden, spleen, liver and diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB+Gem 87 150 20.00 1.46 16.44 0 Moderate tumor burden, spleen, liver and diaphragm invasionAverage: 2.68


0 18 37 89

Time (days)

Tumor implantation

Treatment windows

Gem or PBS

Control Exo or iExo

A

B Survival (Days) Pre-treatment End-point Observations

Treatment PTS PB BW(g) TW(g) BW(g) Lung Metastases Necropsy notes

CB 46 116 19.50 4.14 25.50 6Large primary tumor, secondary tumor, peritoneal, diaphragm, stomach, spleen and liver invasion, mesenteric lymph node metastasis,

macroscopic lung nodules

CB 37 107 19.00 5.04 24.95 >100 Moderate tumor, spleen, liver, diaphragm and stomach invasion, macroscopic lung nodules

CB 47 117 20.00 1.36 16.90 0 Small tumor, bleeding, diaphragm invasion, mesenteric lymph node metastasis

CB 37 107 20.00 5.35 22.56 34 Moderate tumor, paraaortic lymph node metastasis, diaphragm and spleen invasion, possible stomach invasion, macroscopic lung nodules.

CB 42 113 19.50 1.80 25.00 14Moderate tumor, bleeding, mesenteric lymnph node metastases, diaphragm invasion, liver invasion, spleen invasion, macroscopic lung

nodules.

CB 37 107 19.00 2.73 18.71 9 Moderate tumor, spleen, lymph node and diaphragm invasion, macroscopic lung nodules

CB 53 124 20.00 3.46 26.04 3Large primary tumor, secondary tumor, peritoneal, diaphragm, spleen and liver invasion, mesenteric lymph node metastases, macroscopic

lung nodules

Average: 3.41

MSCs Ctrl Exo 42 113 19.00 5.43 29.60 9Large primary tumor, bleeding, ascites, peritoneal, diaphragm, stomach and kidney invasion, lymph node metastases, macroscopic lung

nodules

MSCs Ctrl Exo 42 113 20.00 1.63 22.00 2 Moderate tumor, diaphragm, spleen, peritoneal and lymph node invasion, macroscopic lung nodules

MSCs Ctrl Exo 60 131 19.5 7.12 27.8 >100Large primary tumor, bleeding, ascites, liver, diaphragm, stomach, ovary and spleen invasion, mesenteric lymph noode metastases,


MSCs Ctrl Exo 39 109 20.00 2.02 22.16 >100Moderate tumor, diaphragm invasion, liver invasion, spleen invasion, peritoneal invasion, Mesenteric lymph node invasion, macroscopic lung

nodules

MSCs Ctrl Exo 41 112 19.50 4.18 23.37 32 Large primary tumor, bleeding, diaphragm invasion, stomach invasion, kidney invasion, spleen invasion, macroscopic lung nodules

MSCs Ctrl Exo 52 123 20.00 3.72 25.18 3 Large primary tumor, ascites, bleeding, diaphragm, stomach, liver, kidney and spleen invasion, lymph node metastases, paraaortic lymph node metastases, hydronephrosis, ovarian metastases, macroscopic lung nodules

MSCs Ctrl Exo 39 109 20.00 2.55 22.50 16Moderate tumor, bleeding, ascites, diaphragm, stomach, liver, spleen and peritoneal invasion, mesentereic lymph node invasion,


Average: 3.81

Gemcitabine 54 125 20.00 3.08 25.96 19Large primary tumor, bleeding, ascites, spleen, liver, stomach, diaphragm, peritoneal, ovarian and left kidney invasion, pleural bleeding,


Gemcitabine 56 127 20.00 3.49 25.38 >100Large primary tumor, bleeding, ascites, diaphragm, stomach, ovary, spleen and small instestine invasion, posible liver invasion,


Gemcitabine 67 138 21.00 2.94 26.80 7Moderate tumor divided in two masses connected with necrotic core, very discolored liver (potential metastases), ovary and diaphragm

invasion, macroscopic lung nodules.

Gemcitabine 89** 160 20.00 1.03 21.90 0 Ascites, small tumor, possible spleen invasion and inflammation, necrotic liver

Gemcitabine 54 125 19.50 2.46 22.47 33Moderate tumor, bleeding, ascites, spleen, ovary and diaphragm invasion, hepatic lymph node metastases, paraaortic mesenteric lymph

node metastases, macroscopic lung nodules

Gemcitabine 67 138 20.00 2.49 27.70 2 Moderate tumor divided in two mass connected, very discolored liver (potential metastases), macroscopic lung nodules

Gemcitabine 89** 160 19.50 1.63 25.60 0Moderate tumor, bleeding, ascites, primaric tumor attached to the abdominal wall and connected to the pancreas. Possible spleen invasion

and inflammation, peripancreatic lymph node metastases, diaphragm invasion

Average: 2.45MSCs siKrasG12D-2 iExo CB 89** 160 20.00 2.58 23.60 0 Amall primary tumor, some ascites, peritoneal secondary tumor, diaprahgm invasion

MSCs siKrasG12D-2 iExo CB 66 137 20.00 4.18 25.58 5Large primary tumor with necrotic core, small secondary tumors (2), bleeding, ascites, lymph node metastases, spleen, liver, stomach and

kidney invasion, diaphragm metastases, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB 43 114 20.00 0.81 16.81 0 Moderate tumor, tumor with necrotic core (hole), bleeding, ascites, diaphragm invasion

MSCs siKrasG12D-2 iExo CB 56 127 20.00 5.06 25.90 34Large primary tumor, tumor with necrotic core (hole), bleeding, ascites, mesenteric lymph node and colon invasion, macroscopic lung

nodules

MSCs siKrasG12D-2 iExo CB 89** 160 20.00 2.4 16.00 29Moderate primary tumor, secondary tumor not connected to the primary tumor, tumor with necrotic core (hole), diapragm invasion,


MSCs siKrasG12D-2 iExo CB 67 138 20.00 7.7 30.21 2 Large primary tumor, peritoneal lymph node and diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB 89** 160 20.00 0.2 23.40 0 Remnant of primary tumor, some ascites, no invasion

Average: 3.28MSCs siKrasG12D-2 iExo CB+Gem 89** 160 20.00 1.80 23.10 0 Small primary tumor, no invasion

MSCs siKrasG12D-2 iExo CB+Gem 89** 160 19.50 0.20 20.10 0 Very small, remnant of tumor, large area of normal pancreas, edema in the capsule, no invasion

MSCs siKrasG12D-2 iExo CB+Gem 89** 160 20.00 2.24 23.30 0 Small primary tumor, area of normal pancreas, edema around the capsule, no invasion

MSCs siKrasG12D-2 iExo CB+Gem 89** 160 20.00 1.70 23.10 7Some ascites, spleen and diaphragm invasion, possible liver invasion, mesenteric lymph node metastases, lymph node metastases around

the stomach, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB+Gem 59 130 20.00 1.52 22.56 20 Small primary tumor, diaphragm invasion, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB+Gem 65 136 20.00 3.25 30.88 9Small primary tumor, bleeding, ascites, spleen, stomach, kidney, ovary and diaphragm invasion, hepatic lymph node metastases, lymph

node metastases, macroscopic lung nodules

MSCs siKrasG12D-2 iExo CB+Gem 89** 160 20.00 3.68 25.00 18Large primary tumor with area of normal pancreas, some ascites, bleeding, subcutaneous edema in the left flank, diaphragm invasion,


Average: 2.06

A

PBS BJ Exo2.0

2.5

3.0

3.5

4.0

4.5A

lbum

in g

/dL

PBS BJ Exo0

50

100

150

Alk

alin

e P

ho

sph

ata

se U

/L

PBS BJ Exo0

50

100

150

200

250

ALT

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200

300

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74K-Mesenteric lymph node-macrophages with yellow pigment

75K-Kidney inflammation, acute multifocal interstitial

75K-Liver inflammation-acute multifocal

74K-Liver hyperplasia bile ducts

69K-Normal Liver

72k-Inflammation gall bladder

69K Normal gall bladder

B

Spleen ThymusBone marrow


A

B

CB liposomes BJ Exo MSC Exo MSC iExo0

5

10

15

20

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(pg

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25

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35

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‡

Lung

Kidney

Intestine

Spleen

Femur

Pancreas/Tumor

Liver

Lung

Kidney

Intestine

Testis

Femur

Liver

Spleen

Pancreas

Supplemental Figure 12A

Control DiR

Non tumor bearing Tumor bearing

Mouse 1 Mouse 2 Control DiR Mouse 1 Mouse 2

B

Kidney

Intestine

Femur

Lung

Brain

Heart

Kidney

Intestine

Femur

Lung

Brain

Heart

Pancreas

Spleen

Liver

Control DiR i.p. i.v.Control DiR i.p. i.v.

Radiant Efficiency

p/sec/cm2 /sr

!W/cm

2

0.5x108

1.5 x 108

Brain

Heart

Lung

Stomach

Bowel

Liver

Kidney

SpleenOvary

Femur

Control DiR 24hrs 48hrsControl DiRC

Radiant Efficiency p/sec/cm

2 /sr!W

/cm2

0.5 x108

2.5 x 108

2x108

Radiant Efficiency p/sec/cm

2 /sr!W

/cm2

6.0 x 108

Control DyR i.p. i.v.Pancreas

0.5 x108

2.5 x 108R

adiant Efficiency p/sec/cm

2 /sr!W

/cm2

1.0 x107

6.0 x 107

Radiant Efficiency

p/sec/cm2 /sr

!W/cm

2

1.0 x108

3.0 x 108

Supplemental Figure 13A

B

Untreated Control Exo

5.1%

8.7%

2%

8.3%

57%

21%

57%

30%

FreshFrozen (3 months)

/Thaw

Untreated Control Exo

FreshFrozen (6 months)

/Thaw

1%

4.3%

75.4%

7.5%

83%

3.4%

5.5%

3.1%

Annexin V Annexin V

Annexin V Annexin V

Annexin V Annexin V

Annexin V Annexin V

Mouse ID Treatment Age: Treatment start Survival:PB BW (g) TW (g)

sr31 siKrasG12D iExo 65 117 13.52 0.28sr96 siKrasG12D iExo 66 112 12.63 1.1sr23 siKrasG12D iExo 69 122 19.66 0.75

sm782 siKrasG12D iExo 75 140 8.17 0.14J44 siKrasG12D iExo 60 150 33.1 1.03J45 siKrasG12D iExo 60 98 16.8 1.44J43 siKrasG12D iExo 60 158 20.9 <0.1

Average 17.83 0.79sr29 Control Exo 64 110 18.73 0.3

sr257 Control Exo 58 92 18.2 1.28sr270 Control Exo 60 69sr271 Control Exo 60 85 14.45 0.94sr475 Control Exo 59 101 16.2 1.45J54 Control Exo 61 113 16.66 1.3

Average 16.85 1.05

End point

N/A

LIVE

/DEA

D™

Fixa

ble

Aqua

Sta

inLI

VE/D

EAD

™Fi

xabl

e Aq

ua S

tain

LIVE

/DEA

D™

Fixa

ble

Aqua

Sta

inLI

VE/D

EAD

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ua S

tain

Treatment Group PBS PBS PBS Exo Exo Exo ExoAnimal ID 69K 70k 71K 72K 73K 74K 75KSpecie mouse mouse mouse mouse mouse mouse mouseStrain/Genotype C57BL/6J C57BL/6J C57BL/6J C57BL/6J C57BL/6J C57BL/6J C57BL/6JTerminal body weight (grams) 37.16 35.04 40.28 35.81 32.52 27.84 29.76Liver weight (grams) 2.02 1.99 1.99 1.97 1.71 1.47 1.52Kidney weight (grams) - Right/Left 0.49 0.45 0.57 0.51 0.46 0.43 0.45Heart weight (grams) 0.17 0.21 0.25 0.15 0.16Spleen weight (grams) 0.08 0.07 0.11 0.08 0.07 0.10 0.10Block No.

OrganMorphologic Diagnosis

Lesion Grade

Lesion Grade

Lesion Grade

Lesion Grade

Lesion Grade

Lesion Grade

Lesion Grade

A Liver A A N N A A AInflammation, acute, mixed, multifocal 1 1 2Inflammation, acute, bile ducts 1 2Hyperplasia, bile ducts 1 1Extramedullary hematopoiesis, (incidental finding) 1 1 2

A Gall Bladder N N N A A A AInflammation, acute, neutrophilic 2 1 1Distended/dilated 3

A Kidneys A A A A A A AInflammation, acute, mixed (neutrophils and macrophages), interstitial, multifocal 2

Chronic Progressive Nephropathy (multifocal hyperplasia of cortical renal tubules; and/or proteinaceous tubular casts in the medullary renal tubules) (incidental age-related change)

1 1 1 1 1 2

Lymphoplasmacytic infiltration, perivascular, arcuate and mid-size blood vessels (incidental age-related change)

1 1 1 2 1 1 2

B Lung A N A N N A N

Inflammation, acute, mixed, perivascular 1 2 2Increased number of alveolar macrophages, diffuse 1

C Heart N N N N N N N

E Brain N A N N A N NMineralization/calcification, multifocal, neuropil, thalamus, (incidental finding)

1 1

M Mesenteric Lymph Node and Mesentery 0 A A N A A 0Hyperplasia, paracortex 2 2 2Hyperplasia, lymphoid follicles 2 1Apoptotic lymphocytes with tangible-body macrophages, paracortex

2 3 2 2

Macrophages with intracytoplasmic pale-yellow pigment/material in the medullary sinuses

1 2 3

Inflammation, acute (infiltration of neutrophils) 2Infiltration of mast cells in the medullary sinuses 2Plasmacytosis (increased number of plasma cells) 2

M Spleen N N A A A A AHyperplasia, lymphocytic, periarteriolar lymphoid sheaths (white pulp)

1 2 1

Plasmacytosis (increased number of plasma cells) 2 1 2 1 2T Bone marrow, femur, knee joint N N N N N N N

U Bone marrow, sternum N N N A N N NPlasmacytosis, focal 1

Comment:Liver: Minimal to mild acute inflammation around the bile ducts in mice #73K, 74K, 75K, and of gall bladder in mice #72K, 73K, 75K, indicate mild injury to cholangiolarepithelium of bile ducts and gall bladder. Mild multifocal acute inflammation of mononuclear cells and neutrophils in the liver of mouse #75K is consistent with mild injury/toxicity to hepatocytes. Kidney: Mild acute multifocal interstitial inflammation suggests mild toxicity to epithelium of renal tubules. Immune System: Minimal to mild plasmacytosis and lymphoid hyperplasia of spleen and mesenteric lymph nodes in mice 71K, 72K, 73K, 74K, 75K indicate mild immune activation of lymphoid system and increased production of antibodies by plasma cells. Presence of macrophages with intracytoplasmic pale-yellow pigment in the sinusoids of mesenteric lymph nodes indicates increased phagocytosis of either foreign material, red cells, lipofuscin or ceroid material. Additionally, the mesenteric lymph node of mouse 74K had mild acute inflammatory reaction suggesting a regional injury from gastrointestinal tract or peritoneum.

LEGENDN = Normal tissue or No significant lesion observedA = Abnormal tissue (Lesion present)0 = No tissue present on the slideP = The lesion is present, but not gradedS = Suspected/Presumptive diagnosis (needs more tests to confirm the Dx.)PMA = Postmortem autolysisI = Inadequate sectionN/A= Not ApplicableGRADING/SCORING OF HISTOLOGICAL LESIONSGrade 0 = no histologic lesion (or normal tissue)Grade 1 = minimal, minor, rare, infrequent, barely noticeable tissue change (lesion affects 1-10% of the tissue)Grade 2 = mild, slight, sporadic, noticeable but not prominent feature (lesion affects 11-20% of tissue)Grade 3 = moderate, frequent, typical, common, prominent tissue change (lesion affects 21-40% of tissue)Grade 4 = marked, extensive, numerous, severe, overwhelming tissue change (lesion affects 41-100% of tissue)

Supplemental Table 1

Documents

Supplemental Figure Legend Supplemental Figure 1 A · Supplemental Figure Legend Supplemental Figure 1 (A) Principal component analysis of gene expression of Panc-1 and BxPC3 cells.All